Immediate early genes and methods of use therefor

ABSTRACT

The present invention provides methods and materials related to immediate early genes. Specifically, the invention provides isolated immediate early gene nucleic acid, cells that contain isolated immediate early gene nucleic acid, substantially pure polypeptides encoded by immediate early gene nucleic acid, and antibodies having specific binding affinity for a polypeptide encoded by immediate early gene nucleic acid. In addition, the invention provides cDNA libraries enriched for immediate early genes cDNAs, isolated nucleic acid derived from such cDNA libraries, and methods for treating conditions related to a deficiency in a neuron&#39;s immediate early gene responsiveness to a stimulus.

RELATED APPLICATIONS

[0001] This application claims priority to U.S. provisional applicationNos. 60/074,518, filed Feb. 12, 1998 and 60/074,135, filed Feb. 6, 1998,both of which are incorporated herein by reference.

STATEMENT AS TO FEDERALLY SPONSORED RESEARCH

[0002] Funding for the work described herein was provided by the federalgovernment, which may have certain rights in the invention.

BACKGROUND

[0003] 1. Technical Field

[0004] The present invention generally relates to gene expression andmore specifically to immediate early genes in the brain and polypeptidesencoded by such immediate early genes.

[0005] 2. Background Information

[0006] An immediate early gene (IEG) is a gene whose expression israpidly increased immediately following a stimulus. For example, genesexpressed by neurons that exhibit a rapid increase in expressionimmediately following neuronal stimulation are neuronal IEGs. Suchneuronal IEGs have been found to encode a wide variety of polypeptidesincluding transcription factors, cytoskeletal polypeptides, growthfactors, and metabolic enzymes as well as polypeptides involved insignal transduction. The identification of neuronal IEGs and thepolypeptides they encode provides important information about thefunction of neurons in, for example, learning, memory, synaptictransmission, tolerance, and neuronal plasticity.

SUMMARY

[0007] The present invention involves methods and materials related toIEGs. Specifically, the invention provides isolated IEG nucleic acidsequences, cells that contain isolated IEG nucleic acid, substantiallypure polypeptides encoded by IEG nucleic acid, and antibodies havingspecific binding affinity for a polypeptide encoded by IEG nucleic acid.In addition, the invention provides cDNA libraries enriched for IEGcDNAs, isolated nucleic acid derived from such cDNA libraries, andmethods for treating conditions related to a deficiency in a neuron'sIEG responsiveness to a stimulus.

[0008] In one aspect, the invention features an isolated nucleic acidhaving at least one adenine base, at least one guanine base, at leastone cytosine base, and at least one thymine or uracil base. The isolatednucleic acid is at least 12 bases in length, and hybridizes to the senseor antisense strand of a second nucleic acid under hybridizationconditions. The second nucleic acid has a sequence as set forth in SEQID NO: 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 12, 13, 14, 15, 16, 17, 18, 19,20, 22, 23, 24, 25, 26, 28, 29, 31, 33, 34, 35, 37, 39, 40, 41, 42, 43,44, 45, 46, 47, 49, 50, 51, 52, 53, 54, 55, 56, 57, 58, 59, or 60. Thehybridization conditions can be moderately or highly stringenthybridization conditions.

[0009] In another embodiment, the invention features an isolated nucleicacid having a nucleic acid sequence that encodes an amino acid sequenceat least five amino acids in length. The amino acid sequence contains atleast three different amino acid residues, and is identical to acontiguous portion of sequence set forth in SEQ ID NO: 11, 21, 27, 30,32, 36, 38, 48, 61, or 62.

[0010] Another embodiment of the invention features an isolated nucleicacid having a nucleic acid sequence at least 60 percent identical to thesequence set forth in SEQ ID NO: 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 12, 13,14, 15, 16, 17, 18, 19, 20, 22, 23, 24, 25, 26, 28, 29, 31, 33, 34, 35,37, 39, 40, 41, 42, 43, 44, 45, 46, 47, 49, 50, 51, 52, 53, 54, 55, 56,57, 58, 59, or 60.

[0011] Another embodiment of the invention features an isolated nucleicacid having a nucleic acid sequence that encodes an amino acid sequenceat least 60 percent identical to the sequence set forth in SEQ ID NO:11, 21, 27, 30, 32, 36, 38, 48, 61, or 62.

[0012] Another embodiment of the invention features an isolated nucleicacid having a nucleic acid sequence as set forth in SEQ ID NO: 1, 2, 3,4, 5, 6, 7, 8, 9, 10, 12, 13, 14, 15, 16, 17, 18, 19, 20, 22, 23, 24,25, 26, 28, 29, 31, 33, 34, 35, 37, 39, 40, 41, 42, 43, 44, 45, 46, 47,49, 50, 51, 52, 53, 54, 55, 56, 57, 58, 59, or 60.

[0013] In another aspect, the invention features a substantially purepolypeptide having an amino acid sequence encoded by a nucleic acidhaving at least one adenine base, at least one guanine base, at leastone cytosine base, and at least one thymine or uracil base. The nucleicacid is at least 12 bases in length, and hybridizes to the sense orantisense strand of a second nucleic acid under hybridizationconditions. The second nucleic acid has a sequence as set forth in SEQID NO: 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 12, 13, 14, 15, 16, 17, 18, 19,20, 22, 23, 24, 25, 26, 28, 29, 31, 33, 34, 35, 37, 39, 40, 41, 42, 43,44, 45, 46, 47, 49, 50, 51, 52, 53, 54, 55, 56, 57, 58, 59, or 60.

[0014] In another embodiment, the invention features a substantiallypure polypeptide having an amino acid sequence as set forth in SEQ IDNO: 11, 21, 27, 30, 32, 36, 38, 48, 61, or 62.

[0015] Another embodiment of the invention features a substantially purepolypeptide having an amino acid sequence at least 60 percent identicalto the sequence set forth in SEQ ID NO: 11, 21, 27, 30, 32, 36, 38, 48,61, or 62.

[0016] Another embodiment of the invention features a substantially purepolypeptide having an amino acid sequence at least five amino acids inlength. The amino acid sequence contains at least three different aminoacid residues, and is identical to a contiguous stretch of sequence setforth in SEQ ID NO: 11, 21, 27, 30, 32, 36, 38, 48, 61, or 62.

[0017] Another aspect of the invention features a host cell (e.g., aeukaryotic or prokaryotic cell) containing an isolated nucleic acidhaving at least one adenine base, at least one guanine base, at leastone cytosine base, and at least one thymine or uracil base. The isolatednucleic acid is at least 12 bases in length, and hybridizes to the senseor antisense strand of a second nucleic acid under hybridizationconditions. The second nucleic acid has a sequence as set forth in SEQID NO: 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 12, 13, 14, 15, 16, 17, 18, 19,20, 22, 23, 24, 25, 26, 28, 29, 31, 33, 34, 35, 37, 39, 40, 41, 42, 43,44, 45, 46, 47, 49, 50, 51, 52, 53, 54, 55, 56, 57, 58, 59, or 60.

[0018] Another aspect of the invention features an antibody (e.g., amonoclonal or polyclonal antibody) having specific binding affinity foran amino acid sequence encoded by a nucleic acid having at least oneadenine base, at least one guanine base, at least one cytosine base, andat least one thymine or uracil base. The nucleic acid is at least 12bases in length, and hybridizes to the sense or antisense strand of asecond nucleic acid under hybridization conditions. The second nucleicacid has a sequence as set forth in SEQ ID NO: 1, 2, 3, 4, 5, 6, 7, 8,9, 10, 12, 13, 14, 15, 16, 17, 18, 19, 20, 22, 23, 24, 25, 26, 28, 29,31, 33, 34, 35, 37, 39, 40, 41, 42, 43, 44, 45, 46, 47, 49, 50, 51, 52,53, 54, 55, 56, 57, 58, 59, or 60.

[0019] Another aspect of the invention features a cDNA library having aplurality of clones with each clone having a cDNA insert. In addition,at least about 15 percent (e.g., at least about 20 or 25 percent) of theclones have cDNA derived from immediate early genes (e.g., immediateearly genes responsive to a maximal electroconvulsive seizure). The cDNAlibrary can be a subtracted cDNA library. For example, the subtractedcDNA library can be the IEG-Reg or IEG-Lg cDNA library.

[0020] Another aspect of the invention features an isolated nucleic acidderived from a cDNA library. The cDNA library has a plurality of cloneswith each clone having a cDNA insert. In addition, at least about 15percent of the clones have cDNA derived from immediate early genes. Theisolated nucleic acid can have a nucleic acid sequence of an immediateearly gene.

[0021] Another aspect of the invention features a method of obtainingimmediate early gene nucleic acid. The method includes providing a cDNAlibrary having a plurality of clones with each clone having a cDNAinsert. In addition, at least about 15 percent of the clones have cDNAderived from immediate early genes. The method also includes contactingat least a portion of the cDNA library with a probe containing at leastone nucleic acid having a nucleic acid sequence derived from animmediate early gene, and selecting a member of the plurality of clonesbased on the hybridization of the at least one nucleic acid to themember under hybridization conditions.

[0022] Another aspect of the invention features a method of treating ananimal (e.g., human) having a deficiency in a neuron's immediate earlygene responsiveness to a stimulus. The method includes administering anucleic acid to the animal such that the effect of the deficiency isminimized. The nucleic acid has at least one adenine base, at least oneguanine base, at least one cytosine base, and at least one thymine oruracil base. In addition, the nucleic acid is at least 12 bases inlength, and hybridizes to the sense or antisense strand of a secondnucleic acid under hybridization conditions. The second nucleic acid hasa sequence as set forth in SEQ ID NO: 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 12,13, 14, 15, 16, 17, 18, 19, 20, 22, 23, 24, 25, 26, 28, 29, 31, 33, 34,35, 37, 39, 40, 41, 42, 43, 44, 45, 46, 47, 49, 50, 51, 52, 53, 54, 55,56, 57, 58, 59, or 60. The deficiency can include a reduced level ofexpression of an immediate early gene. In addition, the stimulus caninfluence learning or memory. For example, the stimulus can include amaximal electroconvulsive seizure.

[0023] In another embodiment, the invention features a method oftreating an animal (e.g., human) having a deficiency in a neuron'simmediate early gene responsiveness to a stimulus. The method includesadministering a therapeutically effective amount of a substantially purepolypeptide to the animal such that the effect of the deficiency isminimized. The polypeptide contains an amino acid sequence encoded by anucleic acid having at least one adenine base, at least one guaninebase, at least one cytosine base, and at least one thymine or uracilbase. The nucleic acid is at least 12 bases in length, and hybridizes tothe sense or antisense strand of a second nucleic acid underhybridization conditions. The second nucleic acid has a sequence as setforth in SEQ ID NO: 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 12, 13, 14, 15, 16,17, 18, 19, 20, 22, 23, 24, 25, 26, 28, 29, 31, 33, 34, 35, 37, 39, 40,41, 42, 43, 44, 45, 46, 47, 49, 50, 51, 52, 53, 54, 55, 56, 57, 58, 59,or 60.

[0024] Another embodiment of the invention features a method of treatingan animal (e.g., human) having a deficiency in a neuron's immediateearly gene responsiveness to a stimulus. The method includesadministering an effective amount of cells to the animal such that theeffect of the deficiency is minimized. The cells contain a nucleic acidhaving at least one adenine base, at least one guanine base, at leastone cytosine base, and at least one thymine or uracil base. In addition,the nucleic acid is at least 12 bases in length, and hybridizes to thesense or antisense strand of a second nucleic acid under hybridizationconditions. The second nucleic acid has a sequence as set forth in SEQID NO: 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 12, 13, 14, 15, 16, 17, 18, 19,20, 22, 23, 24, 25, 26, 28, 29, 31, 33, 34, 35, 37, 39, 40, 41, 42, 43,44, 45, 46, 47, 49, 50, 51, 52, 53, 54, 55, 56, 57, 58, 59, or 60.

[0025] Another embodiment of the invention features a method of treatingan animal (e.g., human) having a deficiency in a neuron's immediateearly gene responsiveness to a stimulus. The method includesadministering a therapeutically effective of antibodies to the animalsuch that the effect of the deficiency is minimized. The antibodies havespecific binding affinity for an amino acid sequence encoded by anucleic acid having at least one adenine base, at least one guaninebase, at least one cytosine base, and at least one thymine or uracilbase. The nucleic acid is at least 12 bases in length, and hybridizes tothe sense or antisense strand of a second nucleic acid underhybridization conditions. The second nucleic acid has a sequence as setforth in SEQ ID NO: 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 12, 13, 14, 15, 16,17, 18, 19, 20, 22, 23, 24, 25, 26, 28, 29, 31, 33, 34, 35, 37, 39, 40,41, 42, 43, 44, 45, 46, 47, 49, 50, 51, 52, 53, 54, 55, 56, 57, 58, 59,or 60. The deficiency can include an elevated level of expression of animmediate early gene.

[0026] Another aspect of the invention features a method of identifyinga compound that modulates immediate early gene expression. The methodincludes contacting a test compound with an immediate early gene nucleicacid, and determining whether the test compound effects the expressionof the immediate early gene nucleic acid. The presence of an effectindicates that the test compound is a compound that modulates immediateearly gene expression. The immediate early gene nucleic acid can containa nucleic acid sequence as set forth in SEQ ID NO: 1, 2, 3, 4, 5, 6, 7,8, 9, 10, 12, 13, 14, 15, 16, 17, 18, 19, 20, 22, 23, 24, 25, 26, 28,29, 31, 33, 34, 35, 37, 39, 40, 41, 42, 43, 44, 45, 46, 47, 49, 50, 51,52, 53, 54, 55, 56, 57, 58, 59, or 60. The effect can be a reduction orincrease in the expression of the immediate early gene nucleic acid.

[0027] In another embodiment, the invention features a method ofidentifying a compound that modulates immediate early gene polypeptideactivity. The method includes contacting a test compound with animmediate early gene polypeptide, and determining whether the testcompound effects the activity of the immediate early gene polypeptide.The presence of an effect indicates that the test compound is a compoundthat modulates immediate early gene polypeptide activity. The immediateearly gene polypeptide can contain an amino acid sequence encoded by anucleic acid having at least one adenine base, at least one guaninebase, at least one cytosine base, and at least one thymine or uracilbase. The nucleic acid is at least 12 bases in length, and hybridizes tothe sense or antisense strand of a second nucleic acid underhybridization conditions. The second nucleic acid has a sequence as setforth in SEQ ID NO: 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 12, 13, 14, 15, 16,17, 18, 19, 20, 22, 23, 24, 25, 26, 28, 29, 31, 33, 34, 35, 37, 39, 40,41, 42, 43, 44, 45, 46, 47, 49, 50, 51, 52, 53, 54, 55, 56, 57, 58, 59,or 60. Alternatively, the immediate early gene polypeptide can containan amino acid sequence as set forth in SEQ ID NO: 11, 21, 27, 30, 32,36, 38, 48, 61, or 62. The effect can be a reduction or increase in theactivity of the immediate early gene polypeptide.

[0028] Unless otherwise defined, all technical and scientific terms usedherein have the same meaning as commonly understood by one of ordinaryskill in the art to which this invention pertains. Although methods andmaterials similar or equivalent to those described herein can be used inthe practice or testing of the present invention, suitable methods andmaterials are described below. All publications, patent applications,patents, and other references mentioned herein are incorporated byreference in their entirety. In case of conflict, the presentspecification, including definitions, will control. In addition, thematerials, methods, and examples are illustrative only and not intendedto be limiting.

[0029] Other features and advantages of the invention will be apparentfrom the following detailed description, and from the claims.

DETAILED DESCRIPTION

[0030] The present invention provides methods and materials related toIEGs. Specifically, the invention provides isolated IEG nucleic acid,cells that contain isolated IEG nucleic acid, substantially purepolypeptides encoded by IEG nucleic acid, and antibodies having specificbinding affinity for a polypeptide encoded by IEG nucleic acid. Inaddition, the invention provides cDNA libraries enriched for IEG cDNAs,isolated nucleic acid derived from such cDNA libraries, and methods fortreating conditions related to a deficiency in a neuron's IEGresponsiveness to a stimulus.

[0031] The present invention is based on the discovery of nucleic acidclones for many different neuronal IEGs. Specifically, nucleic acidclones for different neuronal IEGs were isolated and identified based onthe ability of each IEG to rapidly increase expression upon seizureinduction by a maximal electroconvulsive seizure (MECS) method (Cole etal., J. Neurochem. 55:1920-1927 (1990)). It is important to note thatMECS induction can be considered a model to study long-term plasticityrelevant to learning and memory since it is known that a single MECS canproduce extremely robust and long lived potentiation of synapticcontacts in the hippocampus and block spatial learning (Barnes et al.,J. Neurosci. 14:5793-5806 (1994)). Thus, MECS-responsive IEGs caninfluence neuronal activities involved in brain functions such aslearning and memory. Moreover, the isolation and identification of IEGnucleic acid not only provides research scientists with informationabout neuronal activity and gene regulation but also provides methodsand materials that can be used to manipulate brain function.

[0032] Each isolated IEG nucleic acid described herein can be used toproduce a polypeptide. In addition, each IEG nucleic acid can be used toidentify cells that are responsive to MECS induction. For example, anIEG nucleic acid can be labeled and used as a probe for in situhybridization analysis. Clearly, having the ability to identifyMECS-responsive cells provides one with the ability to isolate ormonitor specific brain regions that are involved in learning. Further,any of the isolated partial IEG nucleic acid sequences can be used toobtain a full-length clone that encodes an IEG polypeptide. For example,a fragment from an isolated IEG nucleic acid can be radioactivelylabeled and used to screen a library such that a full-length clone isobtained.

[0033] Cells containing isolated IEG nucleic acid can be used tomaintain or propagate the isolated IEG nucleic acid. In addition, suchcells can be used to produce large quantities of polypeptides that areencoded by isolated IEG nucleic acid. Further, cells containing isolatedIEG nucleic acid can be used to generate virus particles containing theisolated IEG nucleic acid. Such virus particles can be used in vitro orin vivo to provide other cells with the isolated IEG nucleic acid. Thepolypeptides encoded by IEG nucleic acid can be used as immunogens toproduce antibodies. Such antibodies can be used to identifyMECS-responsive cells, monitor the level of polypeptide expressionfollowing MECS induction, and isolate polypeptides directly from animaltissue.

[0034] cDNA libraries enriched for IEG cDNAs can be used to isolatenovel IEG cDNA. Clearly, the isolation of novel IEG cDNAs is importantto further the understanding of brain function. In addition, isolatednucleic acid derived from such cDNA libraries can be used to producepolypeptides as well as identify cells that are responsive to a stimulussuch as MECS induction.

[0035] It is important to note that isolated IEG nucleic acid, cellscontaining isolated IEG nucleic acid, substantially pure IEGpolypeptides, and anti-IEG polypeptide antibodies can be used to treatconditions associated with a deficiency in a neuron's ability to expressIEGs in response to a stimulus such as MECS. A condition associated witha deficiency in a neuron's IEG responsiveness to a stimulus is anyphysiological condition characterized as having a lack of a normal levelof responsiveness. For example, when a deficiency in a neuron'sresponsiveness to MECS is characterized as a non- or under-expression ofa particular IEG polypeptide by that neuron, the organism having thecondition can be treated with isolated IEG nucleic acid, cellscontaining isolated IEG nucleic acid, or substantially pure IEGpolypeptides such that the effect of the deficiency is minimized.Alternatively, when a deficiency in a neuron's responsiveness to MECS ischaracterized as an over-expression of a particular IEG polypeptide bythat neuron, the organism having the condition can be treated withanti-IEG polypeptide antibodies or the anti-sense strand of an isolatedIEG nucleic acid such that the effect of the deficiency is minimized.

[0036] In addition, isolated IEG nucleic acid, cells containing isolatedIEG nucleic acid, substantially pure IEG polypeptides, and anti-IEGpolypeptide antibodies can be used to identify pharmaceutical compoundsthat can be used to treat diseases such as epilepsy, age-dependentmemory decline, stroke, and drug addiction. For example, a compound thatmodulates IEG nucleic acid expression or IEG polypeptide activity can beidentified by contacting a test compound with either the IEG nucleicacid or polypeptide, and determining whether the test compound effectsexpression or activity.

[0037] The term “nucleic acid” as used herein encompasses both RNA andDNA, including cDNA, genomic DNA, and synthetic (e.g., chemicallysynthesized) DNA. The nucleic acid can be double-stranded orsingle-stranded. Where single-stranded, the nucleic acid can be thesense strand or the antisense strand. In addition, nucleic acid can becircular or linear.

[0038] The term “isolated” as used herein with reference to nucleic acidrefers to a naturally-occurring nucleic acid that is not immediatelycontiguous with both of the sequences with which it is immediatelycontiguous (one on the 5′ end and one on the 3′ end) in thenaturally-occurring genome of the organism from which it is derived. Forexample, an isolated nucleic acid can be, without limitation, arecombinant DNA molecule of any length, provided one of the nucleic acidsequences normally found immediately flanking that recombinant DNAmolecule in a naturally-occurring genome is removed or absent. Thus, anisolated nucleic acid includes, without limitation, a recombinant DNAthat exists as a separate molecule (e.g., a cDNA or a genomic DNAfragment produced by PCR or restriction endonuclease treatment)independent of other sequences as well as recombinant DNA that isincorporated into a vector, an autonomously replicating plasmid, a virus(e.g., a retrovirus, adenovirus, or herpes virus), or into the genomicDNA of a prokaryote or eukaryote. In addition, an isolated nucleic acidcan include a recombinant DNA molecule that is part of a hybrid orfusion nucleic acid sequence.

[0039] The term “isolated” also includes any non-naturally-occurringnucleic acid since non-naturally-occurring nucleic acid sequences arenot found in nature and do not have immediately contiguous sequences ina naturally-occurring genome. For example, non-naturally-occurringnucleic acid such as an engineered nucleic acid is considered to beisolated nucleic acid. Engineered nucleic acid can be made using commonmolecular cloning or chemical nucleic acid synthesis techniques.Isolated non-naturally-occurring nucleic acid can be independent ofother sequences, or incorporated into a vector, an autonomouslyreplicating plasmid, a virus (e.g., a retrovirus, adenovirus, or herpesvirus), or the genomic DNA of a prokaryote or eukaryote. In addition, anon-naturally-occurring nucleic acid can include a nucleic acid moleculethat is part of a hybrid or fusion nucleic acid sequence.

[0040] It will be apparent to those of skill in the art that a nucleicacid existing among hundreds to millions of other nucleic acid moleculeswithin, for example, cDNA or genomic libraries, or gel slices containinga genomic DNA restriction digest is not to be considered an isolatednucleic acid.

[0041] Any isolated nucleic acid having a nucleic acid sequence as setforth in SEQ ID NO: 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 12, 13, 14, 15, 16,17, 18, 19, 20, 22, 23, 24, 25, 26, 28, 29, 31, 33, 34, 35, 37, 39, 40,41, 42, 43, 44, 45, 46, 47, 49, 50, 51, 52, 53, 54, 55, 56, 57, 58, 59,or 60 is within the scope of the invention. For convenience, thesenucleic acid sequences will be referred to collectively as the IEGnucleic acid group. In addition, any isolated nucleic acid having anucleic acid sequence at least about 60 percent identical (e.g., atleast about 65, 70, 75, 80, 85, 90, 95, or 99 percent identical) to asequence set forth in the IEG nucleic acid group is within the scope ofthe invention. For the purpose of this invention, the percent identitybetween a sequence set forth in the IEG nucleic acid group (designated atemplate sequence) and any other nucleic acid sequence is calculated asfollows. First, the two nucleic acid sequences are aligned using theMEGALIGN® (DNASTAR, Madison, Wis., 1997) sequence alignment softwarefollowing the Jotun Heim algorithm with the default settings. Second,the number of matched positions between the two aligned nucleic acidsequences is determined. A matched position refers to a position inwhich identical bases occur at the same position as aligned by theMEGALIGN® sequence alignment software. Third, the number of matchedpositions is divided by the total number of bases in the templatesequence, and the resulting value multiplied by 100 to obtain thepercent identity. If the obtained percent identity is greater than orequal to about 60 percent for a particular nucleic acid sequence, thenthat particular nucleic acid sequence is a nucleic acid sequence atleast about 60 percent identical to a sequence set forth in the IEGnucleic acid group.

[0042] Any isolated nucleic acid having a nucleic acid sequence thatencodes an amino acid sequence at least about 60 percent identical(e.g., at least about 65, 70, 75, 80, 85, 90, 95, or 99 percentidentical) to the sequence set forth in SEQ ID NO: 11, 21, 27, 30, 32,36, 38, 48, 61, or 62 is within the scope of the invention. Forconvenience, the amino acid sequences set forth in SEQ ID NO: 11, 21,27, 30, 32, 36, 38, 48, 61, and 62 will be referred to collectively asthe IEG amino acid group. For the purpose of this invention, the percentidentity between a sequence set forth in the IEG amino acid group(designated a template sequence) and any other amino acid sequence iscalculated as follows. First, the two amino acid sequences are alignedusing the MEGALIGN® (DNASTAR, Madison, Wis., 1997) sequence alignmentsoftware following the Jotun Heim algorithm with the default settings.Second, the number of matched positions between the two aligned aminoacid sequences is determined. A matched position refers to a position inwhich identical residues occur at the same position as aligned by theMEGALIGN® sequence alignment software. Third, the number of matchedpositions is divided by the total number of amino acid residues in thetemplate sequence, and the resulting value multiplied by 100 to obtainthe percent identity. If the obtained percent identity is greater thanor equal to about 60 percent for a particular amino acid sequence, thenthat particular amino acid sequence is an amino acid sequence at leastabout 60 percent identical to a sequence set forth in the IEG amino acidgroup.

[0043] Any isolated nucleic acid having a nucleic acid sequence thatencodes an amino acid sequence at least five amino acids in length alsois within the scope of the invention provided the encoded amino acidsequence has at least three different amino acid residues, and isidentical to a contiguous portion of sequence set forth in a sequencewithin the IEG amino acid group.

[0044] Further, any isolated nucleic acid having at least one adeninebase, at least one guanine base, at least one cytosine base, and atleast one thymine or uracil base is within the scope of the inventionprovided the isolated nucleic acid is at least about 12 bases in length(e.g., at least about 14, 15, 16, 17, 18, 19, 20, 25, 30, 40, 50, or 60bases in length), and hybridizes, under hybridization conditions, to thesense or antisense strand of a nucleic acid having a sequence as setforth in the IEG nucleic acid group. The hybridization conditions can bemoderately or highly stringent hybridization conditions.

[0045] For the purpose of this invention, moderately stringenthybridization conditions mean the hybridization is performed at about42° C. in a hybridization solution containing 25 mM KPO₄ (pH7.4), 5×SSC,5× Denharts solution, 50 μg/ml denatured, sonicated salmon sperm DNA,50% formamide, 10% Dextran sulfate, and 1-15 ng/ml probe (>5×10⁷cpm/μg), while the washes are performed at about 50° C. with a washsolution containing 2×SSC and 0.1% SDS.

[0046] Highly stringent hybridization conditions mean the hybridizationis performed at about 42° C. in a hybridization solution containing 25mM KPO₄ (pH7.4), 5×SSC, 5× Denharts solution, 50 μg/ml denatured,sonicated salmon sperm DNA, 50% formamide, 10% Dextran sulfate, and 1-15ng/ml probe (>5×10⁷ cpm/μg), while the washes are performed at about 65°C. with a wash solution containing 0.2×SSC and 0.1% SDS.

[0047] Nucleic acid within the scope of the invention can be identifiedand obtained using any method including, without limitation, commonmolecular cloning and chemical nucleic acid synthesis techniques. Forexample, PCR can be used to obtain a nucleic acid having a nucleic acidsequence at least about 60 percent identical (e.g., at least about 65,70, 75, 80, 85, 90, 95, or 99 percent identical) to a sequence set forthin the IEG nucleic acid group. PCR refers to a procedure or technique inwhich target nucleic acid is amplified in a manner similar to thatdescribed in U.S. Pat. No. 4,683,195, and subsequent modifications ofthe procedure described therein. Generally, sequence information fromthe ends of the region of interest or beyond are used to designoligonucleotide primers that are identical or similar in sequence toopposite strands of a potential template to be amplified. Using PCR, anucleic acid sequence can be amplified from RNA or DNA. For example, anucleic acid sequence can be isolated by PCR amplification from totalcellular RNA, total genomic DNA, and cDNA as well as from bacteriophagesequences, plasmid sequences, viral sequences, and the like. When usingRNA as a source of template, reverse transcriptase can be used tosynthesize complimentary DNA strands.

[0048] Nucleic acid within the scope of the invention also can beobtained by mutagenesis. For example, a nucleic acid sequence set forthin the IEG nucleic acid group can be mutated using common molecularcloning techniques (e.g., site-directed mutageneses). Possible mutationsinclude, without limitation, deletions, insertions, and basesubstitutions, as well as combinations of deletions, insertions, andbase substitutions.

[0049] In addition, nucleic acid and amino acid databases (e.g.,GenBank®) can be used to identify and obtain a nucleic acid within thescope of the invention. For example, any nucleic acid sequence havingsome homology to a sequence set forth in the IEG nucleic acid group, orany amino acid sequence having some homology to a sequence set forth inthe IEG amino acid group can be used as a query to search GenBank®.

[0050] Further, nucleic acid hybridization techniques can be used toidentify and obtain a nucleic acid within the scope of the invention.Briefly, any nucleic acid having some homology to a sequence set forthin the IEG nucleic acid group, or fragment thereof, can be used as aprobe to identify a similar nucleic acid by hybridization underconditions of moderate to high stringency. Such similar nucleic acidthen can be isolated, sequenced, and analyzed to determine whether theyare within the scope of the invention as described herein.

[0051] Hybridization can be done by Southern or Northern analysis toidentify a DNA or RNA sequence, respectively, that hybridizes to aprobe. The probe can be labeled with a radioisotope such as ³²P, anenzyme, digoxygenin, or by biotinylation. The DNA or RNA to be analyzedcan be electrophoretically separated on an agarose or polyacrylamidegel, transferred to nitrocellulose, nylon, or other suitable membrane,and hybridized with the probe using standard techniques well known inthe art such as those described in sections 7.39-7.52 of Sambrook etal., (1989) Molecular Cloning, second edition, Cold Spring harborLaboratory, Plainview, N.Y. Typically, a probe is at least about 20nucleotides in length. For example, a probe corresponding to a 20nucleotide sequence set forth in the IEG amino acid group. can be usedto identify a nucleic acid identical to or similar to a nucleic acidsequence set forth in the IEG nucleic acid group. In addition, probeslonger or shorter than 20 nucleotides can be used.

[0052] Any cell containing an isolated nucleic acid within the scope ofthe invention is itself within the scope of the invention. Thisincludes, without limitation, prokaryotic and eukaryotic cells. It isnoted that cells containing an isolated nucleic acid of the inventionare not required to express the isolated nucleic acid. In addition, theisolated nucleic acid can be integrated into the genome of the cell ormaintained in an episomal state. In other words, cells can be stably ortransiently transfected with an isolated nucleic acid of the invention.

[0053] Any method can be used to introduce an isolated nucleic acid intoa cell. In fact, many methods for introducing nucleic acid into a cell,whether in vivo or in vitro, are well known to those skilled in the art.For example, calcium phosphate precipitation, electroporation, heatshock, lipofection, microinjection, and viral-mediated nucleic acidtransfer are common methods that can be used to introduce nucleic acidinto a cell. In addition, naked DNA can be delivered directly to cellsin vivo as describe elsewhere (U.S. Pat. No. 5,580,859 and U.S. Pat. No.5,589,466 including continuations thereof). Further, nucleic acid can beintroduced into cells by generating transgenic animals.

[0054] Transgenic animals can be aquatic animals (such as fish, sharks,dolphin, and the like), farm animals (such as pigs, goats, sheep, cows,horses, rabbits, and the like), rodents (such as rats, guinea pigs, andmice), non-human primates (such as baboon, monkeys, and chimpanzees),and domestic animals (such as dogs and cats). Several techniques knownin the art can be used to introduce nucleic acid into animals to producethe founder lines of transgenic animals. Such techniques include,without limitation, pronuclear microinjection (U.S. Pat. No. 4,873,191);retrovirus mediated gene transfer into germ lines (Van der Putten etal., Proc. Natl. Acad. Sci., USA, 82:6148-6152 (1985)); genetransfection into embryonic stem cells (Gossler A et al., Proc Natl AcadSci USA 83:9065-9069 (1986)); gene targeting into embryonic stem cells(Thompson et al., Cell, 56:313-321 (1989)); nuclear transfer of somaticnuclei (Schnieke A E et al., Science 278:2130-2133 (1997)); andelectroporation of embryos.

[0055] For a review of techniques that can be used to generate andassess transgenic animals, skilled artisans can consult Gordon (Intl.Rev. Cytol., 115:171-229 (1989)), and may obtain additional guidancefrom, for example: Hogan et al., “Manipulating the Mouse Embryo” ColdSpring Harbor Press, Cold Spring Harbor, N.Y. (1986); Krimpenfort etal., Bio/Technology, 9:844-847 (1991); Palmiter et al., Cell, 41:343-345(1985); Kraemer et al., “Genetic Manipulation of the Early MammalianEmbryo” Cold Spring Harbor Press, Cold Spring Harbor, N.Y. (1985);Hammer et al., Nature, 315:680-683 (1985); Purscel et al., Science,244:1281-1288 (1986); Wagner et al., U.S. Pat. No. 5,175,385; andKrimpenfort et al., U.S. Pat. No. 5,175,384.

[0056] Any method can be used to identify cells that contain an isolatednucleic acid within the scope of the invention. For example, PCR andnucleic acid hybridization techniques such as Northern and Southernanalysis can be used. In some cases, immunohistochemistry andbiochemical techniques can be used to determine if a cell contains aparticular nucleic acid by detecting the expression of a polypeptideencoded by that particular nucleic acid. For example, detection ofpolypeptide X-immunoreactivity after introduction of an isolated nucleicacid containing a cDNA that encodes polypeptide X into a cell that doesnot normally express polypeptide X can indicate that that cell not onlycontains the introduced nucleic acid but also expresses the encodedpolypeptide X from that introduced nucleic acid. In this case, thedetection of any enzymatic activities of polypeptide X also can indicatethat that cell contains the introduced nucleic acid and expresses theencoded polypeptide X from that introduced nucleic acid.

[0057] In addition, any method can be used to force a cell to express anencoded amino acid sequence from a nucleic acid. Such methods are wellknown to those skilled in the art, and include, without limitation,constructing a nucleic acid such that a regulatory element drives theexpression of a nucleic acid sequence that encodes a polypeptide.Typically, regulatory elements are DNA sequences that regulate theexpression of other DNA sequences at the level of transcription. Suchregulatory elements include, without limitation, promoters, enhancers,and the like. Further, any methods can be used to identifying cells thatexpress an amino acid sequence from a nucleic acid. Such methods arewell known to those skilled in the art, and include, without limitation,immunocytochemistry, Northern analysis, and RT-PCR.

[0058] The term “substantially pure” as used herein with reference to apolypeptide means the polypeptide is substantially free of otherpolypeptides, lipids, carbohydrates, and nucleic acid with which it isnaturally associated. Thus, a substantially pure polypeptide is anypolypeptide that is removed from its natural environment and is at least60 percent free, preferably 75 percent free, and most preferably 90percent free from other components with which it is naturallyassociated. Typically, a substantially pure polypeptide will yield asingle major band on a non-reducing polyacrylamide gel.

[0059] Any substantially pure polypeptide having an amino acid sequenceencoded by a nucleic acid within the scope of the invention is itselfwithin the scope of the invention. In addition, any substantially purepolypeptide having an amino acid sequence at least about 60 percent(e.g., at least about 65, 70, 75, 80, 85, 90, 95, or 99 percent)identical to a sequence set forth in the IEG amino acid group is withinthe scope of the invention. The percent identity between particularamino acid sequences is determined as described herein.

[0060] Any method can be used to obtain a substantially purepolypeptide. For example, one skilled in the art can use commonpolypeptide purification techniques such as affinity chromatography andHPLC as well as polypeptide synthesis techniques. In addition, anymaterial can be used as a source to obtain a substantially purepolypeptide. For example, tissue from wild-type or transgenic animalscan be used as a source material. In addition, tissue culture cellsengineered to overexpress a particular polypeptide of interest can beused to obtain substantially pure polypeptide. Further, a polypeptidewithin the scope of the invention can be “engineered” to contain anamino acid sequence that allows the polypeptide to be captured onto anaffinity matrix. For example, a tag such as c-myc, hemagglutinin,polyhistidine, or Flag® tag (Kodak) can be used to aid polypeptidepurification. Such tags can be inserted anywhere within the polypeptideincluding at either the carboxyl or amino termini. Other fusions thatcould be useful include enzymes that aid in the detection of thepolypeptide, such as alkaline phosphatase.

[0061] The term “antibody” as used herein refers to intact antibodies aswell as antibody fragments that retain some ability to selectively bindan epitope. Such fragments include, without limitation, Fab, F(ab′)2,and Fv antibody fragments. The term “epitope” refers to an antigenicdeterminant on an antigen to which the paratope of an antibody binds.Epitopic determinants usually consist of chemically active surfacegroupings of molecules (e.g., amino acid or sugar residues) and usuallyhave specific three dimensional structural characteristics as well asspecific charge characteristics.

[0062] Any antibody having specific binding affinity for an amino acidsequence encoded by a nucleic acid within the scope of the invention isitself within the scope of the invention. Thus, any monoclonal orpolyclonal antibody having specific binding affinity for an amino acidsequence set forth in the IEG amino acid group is within the scope ofthe invention. Such antibodies can be used in immunoassays in liquidphase or bound to a solid phase. For example, the antibodies of theinvention can be used in competitive and non-competitive immunoassays ineither a direct or indirect format. Examples of such immunoassaysinclude the radioimmunoassay (RIA) and the sandwich (immunometric)assay.

[0063] Antibodies within the scope of the invention can be preparedusing any method. For example, any substantially pure polypeptideprovided herein, or fragment thereof, can be used as an immunogen toelicit an immune response in an animal such that specific antibodies areproduced. Thus, an intact full-length polypeptide or fragmentscontaining small peptides can be used as an immunizing antigen. Inaddition, the immunogen used to immunize an animal can be chemicallysynthesized or derived from translated cDNA. Further, the immunogen canbe conjugated to a carrier polypeptide, if desired. Commonly usedcarriers that are chemically coupled to an immunizing polypeptideinclude, without limitation, keyhole limpet hemocyanin (KLH),thyroglobulin, bovine serum albumin (BSA), and tetanus toxoid.

[0064] The preparation of polyclonal antibodies is well-known to thoseskilled in the art. See, e.g., Green et al., Production of PolyclonalAntisera, in IMMUNOCHEMICAL PROTOCOLS (Manson, ed.), pages 1-5 (HumanaPress 1992) and Coligan et al., Production of Polyclonal Antisera inRabbits, Rats, Mice and Hamsters, in CURRENT PROTOCOLS IN IMMUNOLOGY,section 2.4.1 (1992). In addition, those of skill in the art will knowof various techniques common in the immunology arts for purification andconcentration of polyclonal antibodies, as well as monoclonal antibodies(Coligan, et al., Unit 9, Current Protocols in Immunology, WileyInterscience, 1994).

[0065] The preparation of monoclonal antibodies also is well-known tothose skilled in the art. See, e.g., Kohler & Milstein, Nature 256:495(1975); Coligan et al., sections 2.5.1-2.6.7; and Harlow et al.,ANTIBODIES: A LABORATORY MANUAL, page 726 (Cold Spring Harbor Pub.1988). Briefly, monoclonal antibodies can be obtained by injecting micewith a composition comprising an antigen, verifying the presence ofantibody production by analyzing a serum sample, removing the spleen toobtain B lymphocytes, fusing the B lymphocytes with myeloma cells toproduce hybridomas, cloning the hybridomas, selecting positive clonesthat produce antibodies to the antigen, and isolating the antibodiesfrom the hybridoma cultures. Monoclonal antibodies can be isolated andpurified from hybridoma cultures by a variety of well-establishedtechniques. Such isolation techniques include affinity chromatographywith Protein-A Sepharose, size-exclusion chromatography, andion-exchange chromatography. See, e.g., Coligan et al., sections2.7.1-2.7.12 and sections 2.9.1-2.9.3; Barnes et al., Purification ofImmunoglobulin G (IgG), in METHODS IN MOLECULAR BIOLOGY, VOL. 10, pages79-104 (Humana Press 1992).

[0066] In addition, methods of in vitro and in vivo multiplication ofmonoclonal antibodies is well-known to those skilled in the art.Multiplication in vitro can be carried out in suitable culture mediasuch as Dulbecco's Modified Eagle Medium or RPMI 1640 medium, optionallyreplenished by mammalian serum such as fetal calf serum, or traceelements and growth-sustaining supplements such as normal mouseperitoneal exudate cells, spleen cells, and bone marrow macrophages.Production in vitro provides relatively pure antibody preparations andallows scale-up to yield large amounts of the desired antibodies. Largescale hybridoma cultivation can be carried out by homogenous suspensionculture in an airlift reactor, in a continuous stirrer reactor, or inimmobilized or entrapped cell culture. Multiplication in vivo may becarried out by injecting cell clones into mammals histocompatible withthe parent cells (e.g., osyngeneic mice) to cause growth ofantibody-producing tumors. Optionally, the animals are primed with ahydrocarbon, especially oils such as pristane (tetramethylpentadecane)prior to injection. After one to three weeks, the desired monoclonalantibody is recovered from the body fluid of the animal.

[0067] The antibodies within the scope of the invention also can be madeusing non-human primates. General techniques for raising therapeuticallyuseful antibodies in baboons can be found, for example, in Goldenberg etal., International Patent Publication WO 91/11465 (1991) and Losman etal., Int. J. Cancer 46:310 (1990).

[0068] Alternatively, the antibodies can be “humanized” monoclonalantibodies. Humanized monoclonal antibodies are produced by transferringmouse complementarity determining regions (CDRs) from heavy and lightvariable chains of the mouse immunoglobulin into a human variabledomain, and then substituting human residues in the framework regions ofthe murine counterparts. The use of antibody components derived fromhumanized monoclonal antibodies obviates potential problems associatedwith the immunogenicity of murine constant regions when treating humans.General techniques for cloning murine immunoglobulin variable domainsare described, for example, by Orlandi et al., Proc. Nat'l. Acad. Sci.USA 86:3833 (1989). Techniques for producing humanized monoclonalantibodies are described, for example, by Jones et al., Nature 321:522(1986); Riechmann et al., Nature 332:323 (1988); Verhoeyen et al.,Science 239:1534 (1988); Carter et al., Proc. Nat'l. Acad. Sci. USA89:4285 (1992); Sandhu, Crit. Rev. Biotech. 12:437 (1992); and Singer etal., J. Immunol. 150:2844 (1993).

[0069] Antibodies of the present invention also may be derived fromhuman antibody fragments isolated from a combinatorial immunoglobulinlibrary. See, for example, Barbas et al., METHODS: A COMPANION TOMETHODS IN ENZYMOLOGY, VOL. 2, page 119 (1991) and Winter et al., Ann.Rev. Immunol. 12: 433 (1994). Cloning and expression vectors that areuseful for producing a human immunoglobulin phage library can beobtained, for example, from STRATAGENE Cloning Systems (La Jolla,Calif.).

[0070] In addition, antibodies of the present invention may be derivedfrom a human monoclonal antibody. Such antibodies are obtained fromtransgenic mice that have been “engineered” to produce specific humanantibodies in response to antigenic challenge. In this technique,elements of the human heavy and light chain loci are introduced intostrains of mice derived from embryonic stem cell lines that containtargeted disruptions of the endogenous heavy and light chain loci. Thetransgenic mice can synthesize human antibodies specific for humanantigens and can be used to produce human antibody-secreting hybridomas.Methods for obtaining human antibodies from transgenic mice aredescribed by Green et al., Nature Genet. 7:13 (1994); Lonberg et al.,Nature 368:856 (1994); and Taylor et al., Int. Immunol. 6:579 (1994).

[0071] Antibody fragments of the present invention can be prepared byproteolytic hydrolysis of an intact antibody or by the expression of anucleic acid encoding the fragment. Antibody fragments can be obtainedby pepsin or papain digestion of intact antibodies by conventionalmethods. For example, antibody fragments can be produced by enzymaticcleavage of antibodies with pepsin to provide a 5S fragment denotedF(ab′)₂. This fragment can be further cleaved using a thiol reducingagent, and optionally a blocking group for the sulfhydryl groupsresulting from cleavage of disulfide linkages, to produce 3.5S Fab′monovalent fragments. Alternatively, an enzymatic cleavage using pepsinproduces two monovalent Fab′ fragments and an Fc fragment directly.These methods are described, for example, by Goldenberg (U.S. Pat. Nos.4,036,945 and 4,331,647). See also Nisonhoff et al., Arch. Biochem.Biophys. 89:230 (1960); Porter, Biochem. J. 73:119 (1959); Edelman etal., METHODS IN ENZYMOLOGY, VOL. 1, page 422 (Academic Press 1967); andColigan et al. at sections 2.8.1-2.8.10 and 2.10.1-2.10.4.

[0072] Other methods of cleaving antibodies, such as separation of heavychains to form monovalent light-heavy chain fragments, further cleavageof fragments, or other enzymatic, chemical, or genetic techniques mayalso be used provided the fragments retain some ability to selectivelybind its epitope.

[0073] For example, Fv fragments comprise an association of V_(H) andV_(L) chains. This association may be noncovalent, as described in Inbaret al., Proc. Nat'l. Acad. Sci. USA 69:2659 (1972). Alternatively, thevariable chains can be linked by an intermolecular disulfide bond orcross-linked by chemicals such as glutaraldehyde. See, e.g., Sandhu,supra. Preferably, the Fv fragments comprise V_(H) and V_(L) chainsconnected by a peptide linker. These single-chain antigen bindingpolypeptides (sFv) are prepared by constructing a nucleic acid constructencoding the V_(H) and V_(L) domains connected by an oligonucleotide.This nucleic acid construct is inserted into an expression vector, whichis subsequently introduced into a host cell such as E. coli. Therecombinant host cells synthesize a single polypeptide chain with alinker peptide bridging the two V domains. Methods for producing sFvsare described, for example, by Whitlow et al., METHODS: A COMPANION TOMETHODS IN ENZYMOLOGY, VOL. 2, page 97 (1991); Bird et al., Science242:423-426 (1988); Ladner et al., U.S. Pat. No. 4,946,778; Pack et al.,Bio/Technology 11:1271-77 (1993); and Sandhu, supra.

[0074] Another form of an antibody fragment is a peptide coding for asingle CDR. CDR peptides (“minimal recognition units”) can be obtainedby constructing nucleic acid constructs that encode the CDR of anantibody of interest. Such constructs are prepared, for example, byusing PCR to synthesize the variable region from RNA ofantibody-producing cells. See, e.g., Larrick et al., METHODS: ACOMPANION TO METHODS IN ENZYMOLOGY, VOL. 2, page 106 (1991).

[0075] It is also possible to use anti-idiotype technology to producemonoclonal antibodies that mimic an epitope. For example, ananti-idiotypic monoclonal antibody made to a first monoclonal antibodywill have a binding domain in the hypervariable region that is the“image” of the epitope bound by the first monoclonal antibody. Suchanti-idiotypic monoclonal antibodies can be used to inhibit the activityof the polypeptide containing the original epitope.

[0076] The invention also provides cDNA libraries enriched for IEGs. Asdescribed herein, such cDNA libraries contain an increased frequency ofcDNAs derived from IEGs. Specifically, about 15 percent (e.g., about 20or 25 percent) of the cDNA clones within the cDNA libraries providedherein are derived from IEGs.

[0077] A cDNA library within the scope of the invention can be preparedfrom any tissue containing cells that express an IEG (e.g., hippocampustissue). Again, an IEG is a gene whose expression is rapidly increasedimmediately following a stimulus. The stimulus can be electrical orchemical in nature. For example, cells can be treated with electricshock or chemicals such as kainate. Briefly, cDNA libraries are preparedfrom the hippocampus of control animals (e.g., rats) as well as fromanimals that receive a stimulus (e.g., multiple MECS) using, forexample, a phage vector lambda ZAP II (Stratagene). A subtracted libraryis then prepared using in vitro mRNA prepared from a control library andsubsequent solution phase hybridization with cDNA prepared from astimulated library. The control in vitro mRNA can be tagged with biotinto permit its removal from solution using avidin beads (Lanahan et al.,Mol. Cell. Biol. 12:3919-3929 (1992)). cDNA that remains after removalof mRNA/cDNA hybrids can be recloned into, for example, a lambda ZAPIIphage vector. Several rounds of subtraction (e.g., two, three, four, orfive rounds) can be used to increase the frequency of IEGs. Thesubtracted library then can be plated and duplicate phage lifts screenedwith a radiolabeled cDNA probe. Any probe can be used provided itcontains at least one nucleic acid sequence derived from an IEG. Forexample, a probe can be prepared from mRNA obtained from the hippocampusof a stimulated animal. In addition, the mRNA used to make a probe canbe subjected to subtractive hybridization such that IEG sequences areenriched. In general, conventional cDNA libraries contain IEGs at afrequency of <1:30,000 cDNAs. For the cDNA libraries enriched for IEGs,however, about 1 in 5 genes can be induced by a stimulus such as MECS.This represents an about 1000 to 10,000 fold enrichment in brain IEGs.

[0078] An animal (e.g., human) having a deficiency in a neuron's IEGresponsiveness to a stimulus (e.g., a stimulus that influences learningor memory) can be treated using the methods and materials describedherein. A stimulus that influences learning or memory can be a multipleMECS treatment. A deficiency in a neuron's IEG responsiveness to astimulus means the level of IEG responsiveness is not normal. Suchdeficiencies can be identified by stimulating a sample of cells andmeasuring the levels of IEG expression. If the levels are not similar tothe levels normally observed in a similar tissue sample, then there is adeficiency. It is noted that increased IEG expression as well asdecreased IEG expression can be classified as a deficiency provided thelevels are not normal.

[0079] A deficiency in a neuron's IEG responsiveness to a stimulus canbe treated by administering a nucleic acid of the invention to theanimal such that the effect of the deficiency is minimized. Theadministration can be an in vivo, in vitro, or ex vivo administration asdescribed herein. For example, an in vivo administration can involveadministering a viral vector to the hippocampal region of an animal,while an ex vivo administration can involve extracting cells from ananimal, transfecting the cells with the nucleic acid in tissue culture,and then introducing the transfected cells back into the same animal.

[0080] In addition, a deficiency in a neuron's IEG responsiveness to astimulus can be treated by administering a therapeutically effectiveamount cells containing isolated IEG nucleic acid, substantially pureIEG polypeptides, anti-IEG polypeptide antibodies, or combinationsthereof. A therapeutically effective amount is any amount that minimizesthe effect of the deficiency while not causing significant toxicity tothe animal. Such an amount can be determined by assessing the clinicalsymptoms associated with the deficiency before and after administering afixed amount of cells, polypeptides, or antibodies. In addition, theeffective amount administered to an animal can be adjusted according tothe animal's response and desired outcomes. Significant toxicity canvary for each particular patient and depends on multiple factorsincluding, without limitation, the patient's physical and mental state,age, and tolerance to pain. The cells, polypeptides, or antibodies canbe administered to any part of the animal's body including, withoutlimitation, brain, spinal cord, blood stream, muscle tissue, skin,peritoneal cavity, and the like. Thus, these therapeutic agents can beadministered by injection (e.g., intravenous, intraperitoneal,intramuscular, subcutaneous, intracavity, or transdermal injection) orby gradual perfusion over time.

[0081] Preparations for administration can include sterile aqueous ornon-aqueous solutions, suspensions, and emulsions. Examples ofnon-aqueous solvents are propylene glycol, polyethylene glycol,vegetable oils such as olive oil, and injectable organic esters such asethyl oleate. Aqueous carriers include water, alcoholic/aqueoussolutions, emulsions or suspensions, including saline and bufferedmedia. Other vehicles for adminstration include sodium chloridesolution, Ringer's dextrose, dextrose and sodium chloride, lactatedRinger's intravenous vehicles containing fluid and nutrientreplenishers, electrolyte replenishers (such as those based on Ringer'sdextrose), and the like. Preservatives and other additives may also bepresent such as, for example, antimicrobials, anti-oxidants, chelatingagents, and inert gases and the like.

[0082] Further, a deficiency in a neuron's IEG responsiveness to astimulus can be treated by administering a therapeutically effectiveamount of a compound that directly interferes with the translation ofIEG nucleic acid. For example, antisense nucleic acid or ribozymes couldbe used to bind to IEG mRNA or to cleave it. Antisense RNA or DNAmolecules bind specifically with a targeted RNA message, interruptingthe expression of the mRNA product. The antisense binds to the messengerRNA forming a double stranded molecule that cannot be translated by thecell. Typically, an antisense oligonucleotides is about 15-25nucleotides in length. In addition, chemically reactive groups, such asiron-linked ethylenediaminetetraacetic acid (EDTA-Fe), can be attachedto an antisense oligonucleotide, causing cleavage of the mRNA at thesite of hybridization. These and other uses of antisense methods toinhibit the translation of nucleic acid are well known in the art(Marcus-Sakura, Anal. Biochem., 172:289 (1988)).

[0083] An oligonucleotide also can be used to stall transcriptionwinding around double-helical DNA and forming a three-strand helix(Maher, et al., Antisense Res. and Dev., 1:227 (1991) and Helene,Anticancer Drug Design, 6:569 (1991)).

[0084] Ribozymes are RNA molecules possessing the ability tospecifically cleave other single-stranded RNA in a manner analogous toDNA restriction endonucleases. By modifying nucleic acid sequences thatencode ribozymes, it is possible to engineer molecules that recognizespecific nucleotide sequences in an RNA molecule and cleave it (Cech, J.Amer. Med. Assn., 260:3030 (1988)). There are two basic types ofribozymes namely, tetrahymena-type (Hasselhoff, Nature, 334:585 (1988))and “hammerhead”-type. Tetrahymena-type ribozymes recognize sequencesthat are four bases in length, while “hammerhead”-type ribozymesrecognize sequences 11 - 18 bases in length. The longer the recognitionsequence, the greater the likelihood that the sequence will occurexclusively in the target mRNA species. Consequently, “hammerhead”-typeribozymes are preferable to tetrahymena-type ribozymes for inactivatinga specific mRNA species. In addition, 18-based recognition sequences arepreferable to shorter recognition sequences. These and other uses ofantisense methods to inhibit the in vivo translation of nucleic acid arewell known in the art (DeMesmaeker et al., Curr. Opin. Struct. Biol.5:343-355 (1995); Gewirtz et al., Proc. Nat'l. Acad. Sci. USA.,93:3161-3163 (1996); and Stein, Chem. Biol. 3:319-323 (1996)).

[0085] Delivery of nucleic acid, antisense, triplex agents, andribozymes can be achieved using a recombinant expression vector such asa chimeric virus or a colloidal dispersion system. Various viral vectorsthat can be utilized for gene therapy include adenoviruses,herpesviruses, vaccinia viruses, and retroviruses. A retroviral vectorcan be a derivative of a murine or avian retrovirus. Examples ofretroviral vectors in which a single foreign gene can be insertedinclude, but are not limited to: Moloney murine leukemia virus (MoMuLV),Harvey murine sarcoma virus (HaMuSV), murine mammary tumor virus(MuMTV), and Rous Sarcoma Virus (RSV). A number of additional retroviralvectors can incorporate multiple genes. All of these vectors cantransfer or incorporate a gene for a selectable marker so thattransduced cells can be identified and generated. In addition, a nucleicacid sequence of interest along with another nucleic acid sequence thatencodes a ligand for a receptor on a specific target cell can beinserted into a viral vector to produce a vector that is targetspecific. For example, retroviral vectors can be made target specific byinserting a nucleic acid sequence that encodes an antibody that binds aspecific target antigen. Those of skill in the art can readily ascertainwithout undue experimentation specific nucleic acid sequences that canbe inserted into a retroviral genome to allow target specific deliveryof the retroviral vector containing the nucleic acid of the invention.

[0086] A colloidal dispersion system can be used to target the deliveryof the nucleic acid of the invention. Colloidal dispersion systemsinclude macromolecule complexes, nanocapsules, microspheres, beads, andlipid-based systems including oil-in-water emulsions, micelles, mixedmicelles, and liposomes. Liposomes are artificial membrane vesicles thatare useful as delivery vehicles in vitro and in vivo. It has been shownthat large unilamellar vesicles (LUV) that range in size from 0.2-4.0 μmcan encapsulate a substantial percentage of an aqueous buffer containinglarge macromolecules. Thus, nucleic acid, intact virions, polypeptides,and antibodies can be encapsulated within the aqueous interior and bedelivered to cells in a biologically active form (Fraley et al., TrendsBiochem. Sci., 6:77 (1981)). In addition to mammalian cells, liposomeshave been used to deliver nucleic acid to plants, yeast, and bacteria.In order for a liposome to be an efficient nucleic acid transfervehicle, the following characteristics should be present: (1)encapsulation of the nucleic acid of interest at high efficiency whilenot compromising its biological activity; (2) preferential andsubstantial binding to a target cell in comparison to non-target cells;(3) delivery of the aqueous contents of the vesicle to the target cellcytoplasm at high efficiency; and (4) accurate and effective expressionof the nucleic acid (Mannino et al., Biotechniques, 6:682 (1988).

[0087] The composition of a liposome is usually a combination ofphospholipids, particularly high-phase-transition-temperaturephospholipids, usually in combination with steroids, especiallycholesterol. Other phospholipids or other lipids also can be used. Thephysical characteristics of liposomes depend on pH, ionic strength, andthe presence of divalent cations.

[0088] Examples of lipids useful in liposome production includephosphatidyl compounds, such as phosphatidylglycerol,phosphatidylcholine, phosphatidylserine, phosphatidylethanolamine,sphingolipids, cerebrosides, and gangliosides. Particularly useful arediacylphosphatidylglycerols, where the lipid moiety contains from 14-18carbon atoms, particularly from 16-18 carbon atoms, and is saturated.Illustrative phospholipids include egg phosphatidylcholine,dipalmitoylphosphatidylcholine, and distearoylphosphatidylcholine.

[0089] The surface of the targeted delivery system may be modified in avariety of ways. In the case of a liposomal targeted delivery system,lipid groups can be incorporated into the lipid bilayer of the liposomein order to maintain the targeting ligand in stable association with theliposomal bilayer. Various linking groups can be used for joining thelipid chains to the targeting ligand. In general, the compounds bound tothe surface of the targeted delivery system will be ligands andreceptors that allow the targeted delivery system to find and “home in”on the desired cells. A ligand may be any compound of interest that willbind to another compound, such as a receptor or antibody.

[0090] Compounds that modulate IEG expression can be identified bycontacting a test compound with an IEG nucleic acid, and determiningwhether the test compound effects expression. Likewise, compounds thatmodulate IEG polypeptide activity can be identified by contacting a testcompound with an IEG polypeptide, and determining whether the testcompound effects polypeptide activity. Contacting includes in solutionand in solid phase, or in a cell. Any type of compound can be used as atest compound including, without limitation, peptides, peptidomimetics,polypeptides, chemical compounds, and biologic agents. In addition, thetest compound can be a combinatorial library for screening a pluralityof compounds. Compounds identified using the method of the invention canbe further evaluated, detected, cloned, sequenced, and the like, eitherin solution or after binding to a solid support, by any method usuallyapplied to the detection of a specific DNA sequence such as PCR,oligomer restriction (Saiki, et al., Bio/Technology, 3:1008-1012, 1985),allele-specific oligonucleotide (ASO) probe analysis (Conner, et al.,Proc. Nat'l. Acad. Sci. USA, 80:278 (1983), oligonucleotide ligationassays (OLAs; Landegren, et al., Science, 241:1077 (1988), and the like.

[0091] The invention will be further described in the followingexamples, which do not limit the scope of the invention described in theclaims.

EXAMPLES Example 1 Construction of Subtracted cDNA Libraries

[0092] The mRNA used to prepare the cDNA libraries was obtained from thehippocampus of adult rats (male or female). Briefly, the hippocampus wasdissected from naive or stimulated rats, and rapidly frozen in liquidnitrogen. The stimulation protocol used to stimulate the rats was asfollows. Rats were injected intraperitoneally with 50 mg of the proteinsynthesis inhibitor cycloheximide (50 mg/ml stock in 50% ethanol) perkilogram of body weight 15 minutes prior to initiating repetitions ofmaximal electroconvulsive seizure (MECS). MECS was induced by passage ofa constant current signal by means of an ECT unit (Ugo, Basil). Thecurrent signal lasted one second with a frequency of 100 Hz. Each pulselasted 0.5 milliseconds, and the current was 90 milliamperes. Thisstimulus caused brief loss of consciousness and a tonic-clonic seizurelasting 30 seconds to one minute. MECS was administered about every 15minutes for a total of 13 administrations over the course of 2.5 to 3hours. Thirty (30) minutes after the last MECS, the rats were sacrificedby decapitation.

[0093] To collect total RNA, the tissue was homogenized in 4Mguanidinium thiocynate using a polytron and then centrifuged through aCsCl cushion. To isolate polyA⁺ RNA, the resulting total RNA waschromatographed on oligo(dT) columns using a commercial oligo(dT) resinand purification protocol (Fastback, Invitrogen). About 50 naive(control) and 50 stimulated rats were used to generate the polyA⁺ mRNAneeded to make the cDNA libraries and perform the Northern blotanalysis.

[0094] A nonsubtracted cDNA library was made using polyA⁺ RNA isolatedfrom rats subjected to MECS. Briefly, cDNA was synthesized and cloneddirectionally into the Lambda ZAP vector yielding a library containing3.6×10⁶ recombinants. This library was designated the 3 hr MECS/CHXlibrary. Differential screening of the 3 hr MECS/CHX library withcontrol and stimulated rat hippocampal cDNA probes yielded several novelIEGs. Analysis of these IEGs revealed that they were relativelyabundant.

[0095] The 3 hr MECS/CHX library was used as starting material forpreparing a subtracted cDNA library highly enriched for IEGs. Asubtracted cDNA library highly enriched for IEGs can allow for thedetection of lower abundance novel IEGs. To make a subtracted cDNAlibrary, DNA template was prepared from the 3 hr MECS/CHX library asfollows.

[0096] The 3 hr MECS/CHX library was amplified and plated on 15 cm NZCYMagarose plates at a density of about 50,000 phage/plate. A total of1.85×10⁶ phage were plated on a total of 37 plates. The plates wereoverlaid with Suspension Media (SM) and the phage particles eluted byswirling at 4° C. overnight. The lysate was collected, and chloroformadded to a final concentration of 5%. The lysate was clarified bycentrifugation, and the phage containing supernatant collected andstored at 4° C. A 300 ml aliquot of the lysate was treated with RNaseA(final concentration of 1 μg/μl) and DNase I (final concentration of 1μg/μl) for three hours at 37° C. Polyethylene glycol (PEG 6000) wasadded to a concentration of 10%, and NaCl added to a concentration of 1M. After mixing well, the lysate was stored at 4° C. overnight to allowphage particles to precipitate. Phage particles were pelleted bycentrifugation, resuspended in 20 ml of SM, and stored at 4° C. Phageparticles were lysed by adding EDTA to a concentration of 10 mM and SDSto a concentration of 0.2% followed by a 20 minute incubation at 68° C.Polypeptides were removed by two extractions withphenol/chloroform/isoamyl alcohol (50:48:2) followed by two extractionswith chloroform/isoamyl alcohol (24:1). The phage DNA contained within40 ml of lysate was precipitated by adding {fraction (1/10)}th volume of3M NaOAc (pH 5.2) followed by the addition of 2 volumes of 100% ethanol.After mixing, the solution was incubated at −20° C. overnight. DNA waspelleted by centrifugation, resuspended in 10 mM Tris, 1 mM EDTA pH 7.5(TE), and reprecipitated overnight. After this second precipitation, theDNA was pelleted by centrifugation and resuspended in 12 ml of 10 mMTris (pH 7.5), 5 mM EDTA, 300 mM NaCl. To remove residual RNA, RNase A(final concentration of 50 μg/ml) was added followed by incubation at37° C. for 1 hour. To remove RNase A, SDS (final concentration of 0.5%)and then Proteinase K (final concentration of 50 μg/ml) was addedfollowed incubation at 37° C. for 1 hour. The DNA lysate was extractedtwice with phenol/chloroform/isoamyl alcohol (50:48:2) followed by oneextraction with chloroform/isoamyl alcohol (24:1). After thisextraction, the DNA lysate was dialyzed against 12 liters of TE for 2days at 4° C. The 300 ml aliquot of phage lysate yielded 7254 μg ofphage DNA. This phage DNA was then used to prepare in vitro polyA⁺ RNA(cRNA).

[0097] To prepare in vitro cRNA, the phage DNA template was linearizedat the 3′ end of the cDNA insert using the restriction enzyme XhoI.Briefly, 1 mg of phage DNA was digested with 1000 U of XhoI for threehours at 37° C. After the three hour incubation, an additional 1000 U ofXhoI was added and the 37° C. incubation continued an additional threehours. XhoI was removed by adding SDS to 0.5% and Proteinase K to 50μg/ml followed by incubation at 37° C. for one hour. Polypeptides wereremoved by three extractions with phenol/chloroform/isoamyl alcohol(50:48:2) followed by one extraction with chloroform/isoamyl alcohol(24:1). The DNA was precipitated with {fraction (1/10)}th volume 3MNaOAc (pH 5.2) and 2 volumes 100% ethanol. The DNA was pelleted bycentrifugation and resuspended in 500 μl TE (1.58 mg/ml final DNAconcentration).

[0098] This linearized DNA was used as template to prepare in vitro cRNAfrom the sense strand of the cDNA inserts. This cRNA is representativeof the initial in vivo population of RNA in the MECS/cycloheximidetreated rat hippocampus. Forty (40) μg of DNA template was incubatedwith 40 mM Tris (pH 7.5), 6 mM MgCl₂, 2 mM spermidine, 10 mM NaCl, 10 mMDTT, 1 U/μl RNasin, 500 μM ATP, 500 μM CTP, 500 μM GTP, 500 μM UTP, and2 U/μl T3 RNA polymerase in a final volume of 300 μl for two hours at40° C. After two hours, an additional 2 U/μl of T3 RNA polymerase wasadded, and the reaction incubated for an additional two hours at 37° C.for a total time of four hours. The DNA template was removed by addingDNaseI (2 U/μg of template) and incubating the mixture at 37° C. for anhour. Polypeptides were removed by two extractions withphenol/chloroform/isoamyl alcohol (50:48:2) followed by one extractionwith chloroform/isoamyl alcohol (24:1). The cRNA was precipitated at 20°C. with one half volume 7.5 M NH₄OAc and 2 volumes 100% ethanol. ThecRNA was pelleted and resuspended in TE. The cRNA was chromatographed onsephadex G-50 columns (NICK columns; Pharmacia) to remove freenucleotides and the concentration of cRNA determined by UV absorbance at260 A. Thirty (30) μg of DNA template yielded 68.6 μg of cRNA. The cRNAwas either stored frozen at −20° C. or precipitated with {fraction(1/10)}th volume 2 M KOAc (pH 5) and 2 volumes 100% ethanol. The 68.6 μgof cRNA was further purified using oligo(dT) column chromatography toselect polyA⁺ cRNA. The cRNA was bound to oligo(dT) under high saltconditions, rinsed with low salt conditions, and eluted with TE (pH7.5). This eluted cRNA was again passed over an oligo(dT) column underhigh salt conditions, rinsed with low salt conditions, and the polyA⁺cRNA eluted with TE (pH 7.5). The two passes on oligo(dT) celluloseyielded 34.2 μg of polyA⁺ cRNA. This polyA⁺ cRNA was then used astemplate for synthesis of first strand cDNA that was then subtractedagainst control brain and liver polyA⁺ RNA.

[0099] Two cDNA synthesis reactions were performed to prepare firststrand cDNA from the polyA⁺ cRNA. One involved using 2 μg of cRNA with asmall amount of ³²P-dCTP to allow for the analysis of subtractionefficiency, and the other involved using 5 μg of cRNA with noradioactive dNTPs. The radioactive cDNA synthesis reaction was asfollows. First, 2 μl cRNA (1 μg/μl in TE), 1 μl Xho(dT) primer (1.4μg/μl), and 8 μl water was combined, and the mixture was incubated at70° C. for ten minutes, quickly spun, and placed on ice. Second, 1 μlRNasin (40 U/μl), 5 μl 5× Reaction Buffer (BRL), 2.5 μl 0.1M DTT, 1.5 μldNTP mix, and 2 μl ³²P dCTP (3000 Ci/mmole) was added, and the mixturewas incubated at room temperature for ten minutes. The dNTP mixcontained 10 mM of each dATP, dGTP, and dTTP as well as 5 mM of methyldCTP. After incubation, 2 μl of Superscript/MMLV RT mix (1:1) was added,and the mixture (25 μl total volume) was incubated at room temperaturefor five minutes followed by a 90 minute incubation at 40° C. Thenonradioactive cDNA synthesis reaction was as follows. First, 5 μl cRNA(1 μg/μl in TE), 2 μl Xho(dT) primer (1.4 μg/μl), and 3 μl water wascombined, and the mixture was incubated at 70° C. for ten minutes,quickly spun, and placed on ice. Second, 1 μl RNasin (40 U/μl), 5 μl 5×Reaction Buffer (BRL), 2.5 μl 0.1M DTT, and 1.5 μl dNTP mix was added,and the mixture was incubated at room temperature for ten minutes. Afterincubation, 5 μl of Superscript/MMLV RT mix (1:1) was added, and themixture (25 μl total volume) was incubated at room temperature for fiveminutes followed by a 90 minute incubation at 40° C.

[0100] After completion, 3.2 μl of 0.5 M EDTA (pH 8.0) was added to theradioactive reaction, and then the radioactive and nonradioactivereactions were combined. For subtractive hybridizations, it wasnecessary to remove the cRNA template by alkaline hydrolysis. This wasdone by adding 25 μl of TE (pH 7.5) and 5.8 μl of 2 M NaOH. Thisresulted in a 20 mM final concentration of EDTA and a 138 mM finalconcentration of NaOH. The mixture was heated for 30 minutes at 68 to70° C., and then 12.2 μl of 1 M Tris (pH 7.5) and 5.8 μl of 2 N HCl wasadded to neutralize the reaction. The final volume was 100 μl of which 2μl was removed and counted to determine the percent incorporation of³²P-dCTP into cDNA. This analysis revealed that 7000 ng of cRNA wasconverted to 2598 ng of first strand cDNA. This first strand cDNA wassubtracted against adult rat brain and liver polyA⁺ RNA.

[0101] For the subtractive hybridizations, the first strand cDNA waschromatographed on a sephadex G-50 column (NICK, Pharmacia) to removeunincorporated dNTPs, especially the unincorporated ³²P-dCTP in order toallow the efficiency of subtraction to be followed. After the cDNA waseluted from the NICK column, it was mixed with 60 μg of adult rat brainpolyA⁺ RNA that was coupled to biotin (2× Bio RNA). The cDNA and polyA⁺RNA mixture was precipitated by adding {fraction (1/10)}th volume 3MNaOAc (pH 5.2) and 2 volumes 100% ethanol. This mixture then waspelleted and resuspended in 20 μl TE (pH 7.5) and 20 μl 2× SubtractionHybridization Buffer (100 mM Hepes (pH 7.6), 0.4% SDS, 4 mM EDTA, 1 MNaCl). The resuspended cDNA and polyA⁺ RNA mixture was then incubated at95° C. for two minutes, quickly spun, and submerged in a 60° C. waterbath for 48 hours to allow hybrids to form between the cDNA andbiotinylated polyA⁺ RNA (BioRNA).

[0102] The cDNA/BioRNA complexes were removed as follows. First, 40 μl1× Subtraction Hybridization Buffer lacking SDS and 20 μl Strepavidin (1mg/ml) was added, and the resulting mixture incubated at roomtemperature for ten minutes. After incubation, the cDNA/BioRNA complexeswere removed by extraction with phenol/chloroform/isoamyl alcohol. Thephenol phase was back-extracted with 1× Subtraction Hybridization Bufferlacking SDS, and the aqueous phases pooled. Once pooled, 20 μlStrepavidin (1 mg/ml) was added, and the resulting mixture incubated atroom temperature for ten minutes. After incubation, remainingcDNA/BioRNA complexes were removed by extraction withphenol/chloroform/isoamyl alcohol. The phenol phase was back-extractedwith 1× Subtraction Hybridization Buffer lacking SDS, and the aqueousphases pooled. The pooled aqueous phases (about 400 μl) were extractedwith chloroform/isoamyl alcohol. At this point, an aliquot of theaqueous phase was counted to determine the amount of cDNA remaining.Results revealed that 78% of the starting cDNA was removed with 22%remaining (572 ng).

[0103] To perform a second round of subtraction, the aqueous phase(about 400 μl) containing the non-hybridizing first strand cDNA wasmixed with 30 μg of adult rat brain polyA⁺ RNA coupled to biotin and 30μg of adult rat liver polyA⁺ RNA coupled to biotin. The cDNA andbiotinylated polyA⁺ RNA was co-precipitated and hybridized as describedfor the first round. In addition, the cDNA/BioRNA complexes were removedas described above, and the percentage of non-hybridizing cDNA remainingwas determined. Results revealed that two rounds of subtraction removed87.5% of the starting cDNA with 12.5% of the starting cDNA remaining.

[0104] A third round of subtraction similar to the second round wasperformed using the remaining cDNA. Analysis of the remaining cDNArevealed that the three rounds of subtraction had removed 90% of thestarting cDNA leaving 10% of the starting cDNA (255 ng).

[0105] The remaining single stranded cDNA was used to synthesize doublestranded cDNA for the subtracted cDNA library. First, the singlestranded cDNA (300 μl) was alkali treated to remove any remaining RNA asfollows. The final concentration of EDTA was adjusted to 20 mM byaddition of 13 μl of 0.5M EDTA, and then 20 μl of 2M NaOH (120 mM finalconcentration) was added. This mixture was incubated at 68° C. for 30minutes and then neutralized by adding 40 μl 1 M Tris (pH 7.5) and 20 μl2 N HCl. The cDNA was precipitated by adding 10 μl glycogen (10 mg/ml),{fraction (1/10)}th volume 3M NaOAc (pH 5.2), and 2 volumes ethanol. ThecDNA then was pelleted, resuspended in 100 μl of TE (pH 7.5), andpurified on a sephadex G-50 column (NICK, Pharmacia). The purified cDNAwas re-precipitated using glycogen, pelleted, and resuspended in TE (pH7.5) as described. Second, 50 μl resuspended cDNA (single stranded,subtracted cDNA), 20 μl 5× Sequenase Buffer, and 13 μl water wascombined, and the mixture incubated at 65° C. for five minutes, 37° C.for ten minutes, and room temperature for 30 minutes. After incubation,5 μl dNTP mix, 5 μl 0.1 M DTT, 2 μl Sequenase (13 U/μl), and 2 μl Klenow(5 U/μl) was added, and the mixture (100 μl final volume) incubated at37° C. for one hour. The dNTP mix contained 10 mM dATP, 10 mM dCTP, 10mM dGTP, and 10 mM dTTP. The reaction was terminated by adding 3 μl of0.5 M EDTA (pH 8.0) followed by two extractions withphenol/chloroform/isoamyl alcohol and a final extraction withchloroform/isoamyl alcohol. The double stranded cDNA was ethanolprecipitated, pelleted by centrifugation, and resuspended in 86 μl TE(pH 7.5).

[0106] The double stranded cDNA was then restriction digested asfollows. Eighty-six (86) μl cDNA, 10 μl 10× EcoRI Reaction Buffer (NEB),2 μl EcoRI (20 U/μl ), and 2 μl XhoI (20 U/μl) was combined, and themixture (100 μl final volume) incubated at 37° C. for one hour. Afterthis incubation, an additional 2 μl EcoRI (20 U/μl) and 2 μl XhoI (20U/μl) was added, and the mixture again incubated at 37° C. for one hour.After digestion, the reaction was extracted twice withphenol/chloroform/isoamyl alcohol followed by one chloroform/isoamylalcohol extraction. The digested cDNA was precipitated with ethanol,pelleted by centrifugation, and resuspended in 40 μl of 10 mM Tris (pH7.5), 1 mM EDTA, 100 mM NaCl, and 20 μl loading buffer. The cDNA wasdivided into two aliquots, and each aliquot was size-fractionated on a 1ml BioGel A-50 m column. The columns were rinsed with 10 mM Tris (pH7.5), 1 mM EDTA, and 100 mM NaCl, with 50 μl fractions being collected.One column was run to select for only relatively long cDNAs while theother was run to select for all cDNAs. These separate pools were thenextracted twice with phenol/chloroform/isoamyl alcohol followed by onechloroform/isoamyl alcohol extraction. The cDNA was precipitated byadding 5 μl yeast tRNA (1 μg/μl) and 2 volumes of 100% ethanol. The cDNAwas pelleted by centrifugation and directionally cloned into lambdaphage UniZAP as follows. For the regular cDNAs (all sizes), 4 μl water,2 μl 5× Ligase Buffer (BRL), 2 μl UniZAP (500 ng/μl), and 2 μl T4 DNALigase (10 U/μl) was added to the pelleted cDNA, and the mixture (10 μlfinal volume) incubated at 14° C. overnight. For the large cDNAs, 2 μlwater, 1 μl 5× Ligase Buffer (BRL), 1 μl UniZAP (500 ng/μl), and 1 μl T4DNA Ligase (10 U/μl) was added to the pelleted cDNA, and the mixture (5μl final volume) incubated at 14° C. overnight. The ligated cDNA wasthen packaged using packing extracts (Stratagene) and titered onXL1-Blue MRF cells. The subtracted 3 hr MECS/CHX cDNA library containinglarge cDNAs (designated IEG-Lg cDNA library) had 239,000 recombinants,and the subtracted 3 hr MECS/CHX cDNA library containing regular cDNAs(designated IEG-Reg cDNA library) had 4,992,000 recombinants. A portionof each library was rescued as pBluescript plasmid, and the cDNA insertsanalyzed. Of 46 plasmids analyzed from the IEG-Lg cDNA library, allcontained cDNA inserts with the average insert size being 1.36kilobases. Of 44 plasmids analyzed from the IEG-Reg cDNA library, 43contained cDNA inserts with the average insert size being 0.9 kilobases.

[0107] Duplicate southern blots containing cDNA from the 44 plasmidsanalyzed from the IEG-Reg cDNA library were probed with control andstimulated subtracted ³²P-oligolabeled cDNA from rat hippocampus. Elevenof the 44 cDNA inserts gave a clear differential signal that wasstronger with the 3 hour MECS/CHX cDNA probe than with the control cDNAprobe. This result indicates that 1 in 4 of the clones in the IEG-RegcDNA library is derived from an IEG.

Example 2 Preparation of Subtracted cDNA Probes

[0108] Subtracted cDNA was prepared using exactly the same protocoldescribed in example 1 with the exception that rather than in vitro cRNAbeing used as the template for cDNA synthesis, polyA⁺ RNA derived fromcontrol rat hippocampi or rat hippocampi from rats treated with the 3hour MECS protocol was used. After first strand cDNA synthesis, the RNAtemplate was denatured by alkaline hydrolysis, and the free nucleotidesremoved by chromatography on sephadex G-50 (NICK, Pharmacia). The cDNAwas precipitated using {fraction (1/10)}th volume 3M NaOAc (pH 5.2), 2μl glycogen (20 mg/ml), and 2 volumes ethanol, pelleted bycentrifugation, and resuspended in TE (pH 7.5). The final concentrationwas 25 ng/μl. The single strand of cDNA was labeled to high specificactivity (2-4×10⁹ cpm/μg) by oligolabelling (Pharmacia) with ³²P dCTP(3000 Ci/mmole). Free nucleotides were removed by chromotography onsephadex G-50 (NICK column, Pharmacia), and the purified ³²P-labeledsubtracted cDNA used to probe the subtracted cDNA libraries.

Example 3 Screen Subtracted Libraries

[0109] The IEG-Reg and IEG-Lg cDNA libraries were plated on NZCYMagarose plates at a density of 500-800 plaques/plate. Duplicatenitrocellulose filter lifts were prepared from each plate using standardtechniques. The filters were prehybridized overnight at 68° C. in 5×SSPE(pH 7.4), 10% dextran sulfate, 0.2% SDS, 5× Denhardt's Solution, and 50μg/ml boiled, sonicated salmon sperm DNA. The first lift from each platewas then hybridized with 4×10⁶ cpm/ml of the control subtracted cDNAprobe and the second lift with 4×10⁶ cpm/ml of the 3 hour MECSstimulated subtracted cDNA probe. Hybridization was done in freshlyprepared 5×SSPE (pH 7.4), 10% dextran sulfate, 0.2% SDS, 5× Denhardt'sSolution, and 100 μg/ml boiled, sonicated salmon sperm DNA at 68° C. forthree days. Filters were washed twice at room temperature for 30 minutesin 2×SSC/0.2% SDS, twice at 60° C. for two hours in 0.5×SSC/0.2% SDS,and then dried and exposed to X-Ray film for one to seven days. Clonesexhibiting greater hybridization signals with the stimulated cDNA probethan those observed with the control cDNA probe were picked for furtheranalysis.

[0110] The putative neuronal IEGs were analyzed by reverse northernanalysis and northern analysis to confirm that they were truedifferentially hybridizing cDNAs. The nucleotide sequence from the endsof these cDNAs was determined, and those sequences not matching thesequences of known genes were used to obtain full-length cDNAs from cDNAlibraries.

Example 4 Construction of a cDNA Library Enriched for Near Full-LengthIEG cDNAs

[0111] Since the initial isolates for all of the IEGs represented smallcDNAs derived from the 3′ regions of the corresponding RNA, it wasnecessary to rescreen other libraries to obtain full-length or nearfull-length cDNAs. For this purpose, a cDNA library enriched forneuronal IEGs with very long inserts was prepared from 3 hour MECS/CHXpolyA⁺ RNA isolated from rat hippocampi. This RNA was already relativelyenriched for neuronal IEGs since the MECS/CHX stimulus produces a largeinduction of IEG expression. Further, the cDNA was synthesized in thepresence of methylmercuric hydroxide to eliminate RNA secondarystructure allowing for the synthesis of long cDNAs using Superscript IIReverse Transcriptase (BRL).

[0112] The basic protocol used to synthesize cDNA was as follows. First,RNA secondary structure was denatured with methylmercuric hydroxidewhich forms adducts with imino groups of uridine and guanosine in theRNA and disrupts Watson-Crick base pairing. Briefly, 22 μl polyA⁺ RNA(0.5 μg/μl in either 10 mM Tris/1 mM EDTA (pH 7.0) or water) wasincubate at 65° C. for five minutes and then cooled to room temperatureover five minutes. Once cooled, 2.2 μl 100 mM CH₃HgOH (90 μl depc'dwater plus 10 μl 1 M CH₃HgOH) was added, and the mixture incubated atroom temperature for one minute. After incubation, 4.4 μl 700 mM2-mercaptoethanol (190 μl depc'd water plus 10 μl 14 M2-mercaptoethanol) was added, and the mixture (final volume 28.6 μl)incubated at room temperature for five minutes.

[0113] Second, the first strand of cDNA was synthesized as follows. Thevolume of the denatured RNA mixture was adjusted by adding 26.4 μl watersuch that the concentration of RNA was 0.2 μg/μl. In the radioactivereaction, 5 μl (1 μg) polyA⁺ RNA, 2 μl 10× Strand 1 Buffer (Stratagene),1.2 μl Strand 1 dNTP mix (Stratagene), 0.8 μl Xho/dT linker primer (1.4μg/μl), 5 μl water, 3 μl dCTP³² 3000 Ci/mmole (NEN), and 1 μl RNaseBlock (Stratagene) was combined, and the mixture (final volume 18 μl)incubated at room temperature for ten minutes to allow the primer toanneal to the RNA. In the nonradioactive reaction, 25 μl (5 μg) polyA⁺RNA, 5 μl 10× Strand 1 Buffer (Stratagene), 3 μl Strand 1 dNTP mix(Stratagene), 2 μl Xho/dT linker primer (1.4 μg/μl), 9 μl water, and 1μl RNase Block (Stratagene) was combined, and the mixture (final volume45 μl) incubated at room temperature for ten minutes to allow the primerto anneal to the RNA. After the room temperature incubation, 2 μl and 5μl of reverse transcriptase mix (4 μl Superscript II (BRL 200 U/μl) plus1 μl MMLV RT (Stratagene)) was added to the radioactive andnonradioactive reactions, respectively. The reactions then wereincubated at 40° C. for one hour and placed on ice. Two μl of cDNA wasremoved from the radioactive reaction and added to 18 μl T₁₀E₁ and 2 μl0.5M EDTA. Two (2) μl of this mixture then was applied to a PEI strip todetermine the percent incorporation and quantity of cDNA synthesized,while 18 μl was mixed with sample buffer and ran on a gel to assay cDNAquality.

[0114] Third, the second strand of cDNA was synthesized as follows. Boththe radioactive and nonradioactive reactions were kept on ice to prevent“snapback” cDNA synthesis. For the radioactive reaction (18 μl), 10 μl10× Second Strand cDNA Buffer, 3 μl Second Strand dNTP mix, 62.5 μlwater, 1 μl RNaseH (1.5 U/μl), and 5.5 μl DNA Polymerase I (9 U/μl) wasadded, and the mixture (100 μl final volume) incubated at 16° C. for 2.5hours. For the nonradioactive reaction (50 μl), 20 μl 10× Second StrandcDNA Buffer, 6 μl Second Strand dNTP mix, 111 μl water, 2 μl RNaseH (1.5U/μl), and 11 μl DNA Polymerase I (9 U/μl) was added, and the mixture(200 μl final volume) incubated at 16° C. for 2.5 hours. Four (4) μl ofcDNA was removed from the radioactive reaction and added to 18 μl T₁₀E₁and 2 μl 0.5M EDTA. Two μl of this mixture then was applied to a PEIstrip to determine the percent incorporation and quantity of cDNAsynthesized, while 18 μl was mixed with sample buffer and ran on a gelto assay cDNA quality.

[0115] The cDNA from both the radioactive and nonradioactive reactionswere extracted twice with phenol/chloroform/isoamyl alcohol followed byone extraction with chloroform/isoamyl alcohol. After extraction, thecDNA was precipitated with 100% ethanol, pelleted by centrifugation, andresuspended in 39.5 μl water. To blunt the cDNA ends, 5 μl 10× T4 DNAPolymerase Buffer (NEB), 2.5 μl dNTP mix (2.5 mM each dNTP), and 3 μl T4DNA Polymerase (3 U/μl) was added to the 39.5 μl of cDNA, and themixture (50 μl final volume) incubated at 16° C. for 30 minutes. Afterincubation, 350 μl TE (pH 7.5) was added, and the mixture (400 μl finalvolume) extracted twice with phenol/chloroform/isoamyl alcohol followedby one extraction with chloroform/isoamyl alcohol. After extraction, thecDNA was precipitated with 100% ethanol, pelleted by centrifugation, andresuspended in 17 μl water.

[0116] EcoRI/NotI adaptors were ligated to the cDNA, allowing for thequick identification of artifactual cDNAs generated by the ligation oftwo independent cDNAs prior to ligation into the lambda phage vector. Toligate the EcoRI/NotI adaptors to the cDNA, 3 μl 10× Ligase Buffer, 4 μlEcoRI/NotI Adaptors (1 μg/μl), 3 μl 10 mM ATP, and 3 μl T4 DNA Ligase(400 U/μl) was added to the 17 μl cDNA, and the mixture (30 μl finalvolume) incubated at 10° C. overnight. After the overnight incubation, 1μl T4 DNA Ligase and 1 μl 10 mM ATP was added, and the IS mixture (32 μlfinal volume) again incubated at 10° C. overnight. After this secondovernight incubation, 270 μl TE (pH 7.5) was added and the mixtureextracted twice with phenol/chloroform/isoamyl alcohol followed by oneextraction with chloroform/isoamyl alcohol. After extraction, the cDNAwas precipitated with 100% ethanol, pelleted by centrifugation, andresuspended in 30 μl water.

[0117] To kinase the cDNA ends, 4 μl 10× T4 Polynucleotide KinaseBuffer, 4 μl 10 mM ATP, and 2 μl T4 Polynucleotide Kinase (10 U/μl) wasadded to the 30 μl of cDNA, and the mixture (40 μl final volume)incubated at 37° C. for 30 minutes. After incubation, 2 μl T4Polynucleotide Kinase was added, and the mixture (42 μl final volume)incubated at 37° C. for 30 minutes. After this second 30 minuteincubation, 170 μl TE (pH 7.5) was added, and the mixture extractedtwice with phenol/chloroform/isoamyl alcohol followed by one extractionwith chloroform/isoamyl alcohol. After extraction, the cDNA wasprecipitated with 100% ethanol, pelleted by centrifugation, andresuspended in 85 μl water.

[0118] To digest the 3′ cDNA ends with XhoI, 10 μl 10× NEB Buffer #2 and5 μl XhoI (20 U/μl) was added to the 85 μl of cDNA, and the mixture (100μl final volume) incubated at 37° C. for 45 minutes. After incubation, 3μl XhoI (40 U/μl) was added, and the mixture (103 μl final volume) againincubated at 37° C. for 45 minutes. After this second incubation, 120 μlTE (pH 7.5) was added, and the mixture extracted twice withphenol/chloroform/isoamyl alcohol followed by one extraction withchloroform/isoamyl alcohol. After extraction, the cDNA was precipitatedwith 100% ethanol, pelleted by centrifugation, and resuspended in 20 μl10 mM Tris (pH 7.5), 1 mM EDTA, 100 mM NaCl, and 5 μl loading buffer.This resuspended cDNA then was size-fractionated on a 1 ml BioGel A-50mcolumn to select large cDNAs. The column was rinsed with 10 mM Tris (pH7.5), 1 mM EDTA, and 100 mM NaCl. Thirty-six (36) fractions (50μl/fraction) were collected. Aliquots from individual fractions wereelectrophoreses on 1% agarose to identify fractions containing cDNAslonger than 2 kilobases. Such fractions were pooled, and the resultingmixture of pooled fractions was extracted twice withphenol/chloroform/isoamyl alcohol followed by one extraction withchloroform/isoamyl alcohol. After extraction, the cDNA was precipitatedby adding 2 μl glycogen (20 mg/ml) and 2 volumes 100% ethanol, pelletedby centrifugation, and resuspended in 5 μl water.

[0119] To directionally clone the cDNA into UniZAP, 2 μl UniZAP (500ng/μl), 1 μl 10× T4 DNA Ligase Buffer, 1 μl 10 mM ATP, and 1 μl T4 DNALigase (4000 U/μl) was added to the 5 μl of cDNA, and the mixture (10 μlfinal volume) incubated at 12° C. overnight. After incubation, the cDNAwas packaged into phage particles. To package the cDNA, the ligationreaction (10 μl final volume) was divided into two packaging reactionswith each containing 5 μl of ligation reaction together with a packagingextract (Stratagene). This mixture was incubated at 22° C. for 2 hours.After incubation, the two reaction mixtures were pooled and the librarytitered on IL1-Blue MRF cells.

[0120] This 3 hr MECS/CHX library (designated IEG-FL 3 hr MECS/CHX cDNAlibrary) had a titer of 4.4×10⁶ primary phage. The library was amplifiedand used to isolate full length cDNAs derived from novel neuronal IEGs.The relative abundance of near full length neuronal IEG cDNAs in thislibrary was substantially higher than the levels experienced using othercDNA libraries. In a single cDNA library screen, full length cDNAs forfour different novel IEGs were obtained. Three of the four IEG cDNAswere derived from mRNAs of 4 kilobases, while one was derived from anmRNA of 3 kilobases.

[0121] The nucleic acid sequencing of the IEG cDNAs was performed atJohns Hopkins School of Medicine and at Applied Biosciences, Inc., CAusing the Sanger method with fluorescent dye termination.

[0122] Northern blot analysis was performed both to confirm that thecloned cDNAs represent tissue mRNA that is rapidly induced by brainactivation and to assess the size of the mRNA transcript. The latter isessential information for the identification of authentic full lengthclones. Either 20-25 μg of total RNA or 2 μg of polyA⁺ RNA was sized bydenaturing agarose gel chromatography and transferred to nitrocellulose.Blots were then hybridized with [³²P]labeled cDNAs. Labelling was doneusing the random primer method (Pharmacia).

[0123] In addition, in situ hybridization was performed both to confirmthat the cloned cDNAs represent tissue mRNA that is rapidly induced bybrain activation and to confirm that the mRNA was induced in activatedneurons. In situ hybridization was performed as described previously(Andreasson and Worley, Neuroscience 69: 781-796 (1995)).

Example 5 IEG Nucleic Acid

[0124] The following clones were identified as being IEG nucleic acid asdescribed in Example 3. In addition, certain clones were identified bychip-hybridization between PCR fragments generated from rat hippocampusESTs and ³²P-dCTP-labeled cDNA derived from polyA⁺ RNA of rathippocampus from MECS treated animals and controls.

[0125] One IEG nucleic acid clone was designated A003. The first libraryscreen produced a fragment (A003-1-1) of 1.6 kilobases (kb) with a polyAsequence at the 3′-end. A second round of screening was performed usinga probe prepared from the 5′-end of A003-1-1. This screen produced twoclones: A003-1 (2.8 kb) and A003-2 (1.3 kb). The fragments from thesecondary screen were sequenced from both ends. These fragments formed acontig at their 3′-end with the A00-3-1-1 fragment. The following twonucleic acid sequences are within the A003 clone: (SEQ ID NO:1)5′-TTGCAGATCAGCACCTTTTGATGATGCCTGCCCAACAGTGGGTAATGCTNACAGCAAAGCACCACTTTACGCTTTTTAGTTGTGCTGGGTTCATGGCTGGACATACACCAACCAGCCTTGACCCCACAGGAATGCCAAGTTGGCTGGAATGTAACCCAACCTAGTTTCTGCGCTTCGCTCCTCTCCCAGTGCAAGGTGCTAAACACCCACTCACAAGCCTGCTGTCAAGCTGCGACCTTGGGGGCTGGTTAGAAAGGGCTGCCTCCTTCCAGCAATAGAAGTTCATGAATTGAGGCTGGAGATAGGTCAAGACCACTGTGATAACTATAAAGACTGTAGCAGCCACAAAGGAGACCCCCAAATAACTGGAGGCATGGGCACTGACGTACCAGATGAGGTTATGTTTGGAGCTGAAGGCTTGCTCTGTGCTTCTTGGTAGCATCTTTTGTCCTCTTGGGACATGGTTGACCCCATACTGTCCACTGAGCTTGGGAGATGACAGTTGAATAAAAAAAAAAAAAAAAAA-3′ and (SEQ ID NO:2)5′-CGGCTTAATTAACCCTCACTAAAGGGAACAAAAGCTGGAGCTCCACCGCGGTGGCGGCCGCTCTAGAACTAGTGGATCCCCCGGGCTGCAGGATTCTGCGGCCGATTAAGAAGCCTGCTGATGTCCTTAGGCGAGGACATTAACTCCAGTCTCTGACAGACTTTGGACATCCAGAATAAGTTCTTTTTGTATATCAGAGCACAGAGCCCAGCTTTAGCCTCTGATGGACCTCAGGAACCAAGAAGGAGGGACTTCCTTAACATTCTAGAGATGGGACTCTAACTCTAGCTCTTGTGTTAAGCCCTGAAGTCCAGAAAGAAGTAGTTCTTTGACATTCTAGTGCCAAGATCCAGCCTCTAAGAGAACTCTGATGTCTAAAGAAAGTCTTTCATAGTCTAGNCCAGTCACCAGTGAAGCTAAACACCTGAAAACTATTAGATTCTCTGGAGCCAGGAATCCATCTCAAGTCTCTCATAAAGCCCAAATGTCCCAGGAGAAGTTGACAATATAAAGCCGTATCTCGATGGACTTTTGAAGAAGCTCAGAAAAGGAGACCACCTTGGTAGTCTTGATCTAGGACTCTGGCTTGTTTGTCTCCAGGGACGTTTACATGTATAAAAAGAGGGACCTTTCTGATGATTCAGAACTGGGACTCCACCTCCATCCTTTGATGAAAGCTCAAATGTCCAGAAAGAGGGGCCTCTCTGATATTCTAGAGTAGGACCCTCCCTCCAGCCTTTGATGGTGTCCAGATGTCCAGAAAGAGGGGCCTCTCTGATGTTCCAGACCTAGGGCCCTCCCTCCAGCCTTTGATGGTGTCCAGATGTCCAGAAAGAGGGACTTCTCTGATGTTCCAGACCTAAGACTCTAGCTCCAGCCTTTGATGAAGCTCAGATGTTCAGAAAGGGGGGCCTCCATGATGTTCTAGAACCAGGACTCCACCTCTAGCCTTTGATGGTGTCCAGATGTCCAGAAAGAGGGTCTTCCATGATTTCTAGGACCAAGACTTTACCTCCAGCCTTCTATGCCTCCATGTCTCCAGTAAAGCTTAGGTGTCCAGAAAAGAGCATTCTCAATGAATTTATAGAACCAGGACTCTTTCTCCAGCCTTTGATGACGTTCAGATGTTCATAAAGAAGAACTTCCACAATGTACTAAAGCTATGACTCCATCTCCATCCTTTGATGAAAAGGGACTTCCTTCCACTCTGTTCCAGAAGCCTAGCTCCACCTCTAATCTTTGTTGATGTCCAATTATCCAGAAAGAGGGGGCCTTTAGAACAAAGACTGTACTTTTATTCATTGATAAAGCACAGATTCCAGAAGCACAGAAATCTAGAAAGAGGGTCCTCCCTAACACGCTCGAGCTAGAACCCCGGTGCAAGGGTCTGAAACTTAGACACCAGAAGACCGCTTTGTCCTACAACAAGTCTGCATTTTCTAAATCTCCAGGTGGCTGAT:CAGAAGGGTCCAGGAAGGTATGGGG-3′.

[0126] Northern blot analysis using the 3′-end of A003 revealed thepresence of two mRNA transcripts. The more abundant transcript was 2.2kilobases in length, while the less abundant transcript was 4.8kilobases in length. This analysis also revealed that the expression ofA003 mRNA was marginally upregulated in response to the multiple MECStreatment. The multiple MECS treatment involved the induction ofmultiple maximal electroconvulsive seizures followed by the preparationof total RNA from rat hippocampus four hours post-seizure. This multipleMECS treatment was designed to mimic ischemia.

[0127] Another IEG nucleic acid clone was designated A013. The firstlibrary screen produced the clone designated A013-8. The 5′-end ofA013-8 was used as a probe for the second round of screening. Thissecond screening produced two additional clones: A013-4 and A013-26. TheA013-8, A013-4, and A013-26 clones were sequenced using either the genespecific primer used to generate the probe for the second round oflibrary screening, or the T3 and T7 primers. Both A013-4 and A013-26made a contig on their 3′-ends with the A-013-8 clone. In addition, thesequence from the 5′-ends of A013-4 and A013-26 revealed that they fromcontigs between each other. Further, the sequence data from the 5′-endsof A013-4 and A013-26 revealed the presence of an open reading frame ofat least 720 basepairs (bp) Based on the combined length of the obtainedclones, the A013 clone is at least 3.0 kb in length. The following twonucleic acid sequences are within the A013 clone: (SEQ ID NO:3)5′-GGCACGAGATCACTCAGTGTCTTCACTGAACCAAATCGTCATTTTTACAGAGAGATGCAAAGCTTCAGCGAAGACATTTAGCTTTTTTAAAATGTATAATTCCTGTGGCTACATATGCAAGTAGGGTCCCATTATGTTTTTTTTCATTAGTGGAAACTAATCCTTTTGTGCTGTGTTTAATCAGTATTAGCTTTATAGAATTATAAATGTATATTCTACTTCTTGATCAAAGAACGTAGTCGGGTATTGGTTTTAGAAGTTCAAAGTGACACTGTATAGGGCTTTCACGGTTAATGGGATTGTTAGCAAATCTTAAGGACATACAGCCAATGATTATCTGAGGTTACTGGCTAACTGTTTTTCACTGAGTTACTCTGCCTTTTTGACATTTTTATTCTTTGTTTGTCAGAATCCAGAGCTTCAGGAGCCCAAATTTTTTTATWCCGTATATATATATATATATAAATATCCATAAGCCTGGTGGATTTGTATGCAATGCACTGCATCTATGTATTCTGATAGCATCTCATTGATTTTTGTTTGAAATAGAAAGAAAGATAGTATCCCAAATGAGTTATCTTTAACAGAAAGCTGAGTTTAACTTTTATTACCTATATAATAATTGATATTGCCAATTACCATTCTGAATTTCATATAGTATAAGTTAGACATTGCTTAATCCCCTTTTAAATGTATTTACATAGACATGAACACTCAAATTGCTGGATTTTTTAAATATACTGACATAATTTTTTTCATCTGTTACATTCAAGTTAGCTTGTTTAGCCCAGATTTCAGAATAGTAAAGGAGGAAAGGAACCGCATTCCAGGGAAACCTCTGAGGCCAAGTCAGAGTCCAGAACTGTAAACACACAGGCCTGCAAGCCAACATTAGTCGTGAAATCCCTAACACGTCACTGGATTCTCTCTGTCAGCGCAAGTGTCAGCTGCCAAAGAATAGACTTACATGAAGAAGTGCCCACATGCTGGCAGGGGCTGGCCGGCTCCGGCCAGCAGACACTGCTAGATTGTAATATTTAAGGTCGAGTTTCGACCTGTGGTACACAGCTGTGCTGTGCTCAGTCAGCAACCTCAGAACTCTGAAAAAAACATAAAAAAGAAAAAAAAAAAAAAAAAAMATGCASCTGKYTCACTTGTGAATAGTGAATGTAAAGGAAAGAAAGGAAAACCAAAAGCTTGTTCCATCACAGGTATGAGCTGCTATGATTCATGAAGAACATTCCATGGAGTATGTTTTAAAACCTTGTTATATCTGAGAGGCTTTAAAAGCCAACTTAACTGTTTCAGGGCAACCGCGGTACAGACGTGGTCTCTGTGAGACTTCCACCTGACCCAAGTTTTAAGTGGTACGAATGTTGTGCATTTAATGTTAAGGACAGTCTGCAATAATAAGTAAGTAGCCAGCGTGGGTGCCCAGCAGTGCTGAGACCTGGCTGCTCTATTGTACGCTTTGGAAACACAATTTATGCAACAGATGTCCAGATATGATTCTATTTATGGAAAAAGTTTATATGTTTTACAAATGGTTTTACCATCTTATATTAAATGACCTTTTGACAGGTGTGCACTGTTTTGTCTCCAGTGAGCACATACCATGCGGATTTTATATGTACATCAGTAGTGTGAATCCACTGGCACAGTGTGTGTAAATGCCAGATGTGGTGAGATTTTATCTTGTATATGTGATCAGATAAAATAACTCCTGACAGAAACTGTAAGGRAACCCAGCTGAATGGTTTGACCTGGATGRCYKRKRTKGTWTGGTTTATGTTAAATGTATATTCTTTTAATCAATGAATAAAGCATTAAAAAATGGGAAAAAAAAAACTCGTGC-3′ and (SEQID NO:4)5′-TCTGCGGCCGCAGCATCCGGAACAACAGGAACCTCCAGAAGTTTAGTCTTTTTGGAGATATAAGTGTCGTTCAGCAGCAAGGAAGTCTGTCCAGCACATACCTCAGCAGAGTAGACCCTGACGGCAAGAAGATTAAGCAAATTCAGCAGCTGTTTGAAGAGATACTGAGCAATAGTAGGCAACTAAAATGGCTGTCCTGTGGGTTTATGCTGGAAATAGTAACCCCATCATCACTGTCGTCTCTGTCTAACTCCATTGCCAACACCATGGAACACCTGAGTTTACTGGACAACAACATTCCTGGTAACAGCACGCTCATCACCGCAGTCGAACTAGAGCGCTTTGTAAATCTGCGCTCACTTGCCCTGGATTTCTGTGACTTTACAGCTGAGATGGCGAGAGTCCTGACCGAGCAGCAACCATGTGCCTTTGCAGCGACTGTCTCTTCTGGTCCACAATGCTTCAGTGATGCTCAAGTCATTAGACAACATGCCAAACGATGAGCACTGGAAGGCCCTGTCACGAAAGAGCTCCAGCCTCCGGGTCTATCTAATGGCTTTTGATGTTAAAAGTGAAGACATGCTAAAGATTCTGAAACCAGTATACCACTTGAGAAGGGTTCACTTTGGACAGCTACGTCACTTGTGTCTCAAGGGGCTATTGGTTGATCTTATATTCCAGGCAGTATTGACCAAGGTTTCCTYAACCCMWTTTWTATTGATGAATGATATGATTGATACGTCTGGTTTTCCGGATCTTAGTGACAACCGAAATGAAGATCCATTGGTTTTATTGGCATGGCGGTGCACAAAGCTCACTCTTTTGGCAATTCATGGTTACACCGTGTGGGCACACAACCTCATTGCCATTGCTCGTCTTCGTGGCTYTTGACCTAAAAGTGCTTTGGAAGTCACCSRAAGAAAGCATTGATTTTGACCAAGGTGAACTAGCCCGACCAGGAATGTGGRWYCCCGTACATAACCTTTCTTGGAGCAGGTATTCCCTGGGGCCTTGGTCAAGTCTGGCACG-3′.

[0128] Northern blot analysis using a sequence from the A013 clonerevealed the presence of a 3.2 kb mRNA transcript. In addition, thisanalysis revealed that the expression of the A013 mRNA was stronglyupregulated in response to the multiple MECS treatment. Specifically,A013 mRNA expression was induced 8.9 fold by the multiple MECS treatmentas determined from Northern blot data using total RNA from rathippocampus (Table I). TABLE I Fold induction of mRNA expression aftermultiple MECS treatment Probe (rat cDNA) Fold induction (normalized forthe S26) A013 8.9 L094 7.3 L100 17.2 L119 17.8 R113 7.0 R286 2.4

[0129] Another IEG nucleic acid clone was designated A020. The followingnucleic acid sequence is within the A020 clone: (SEQ ID NO:5)5′-TCAAACCNTATCTCGGTCATTCNTTTGATTNATAAGGGATTTKSCCGATKTCCGGCNTATTGGTTAAAAAWTGAGCTGATTTAACAAAAATTTAACGCGAATTTTAACAAAATATTAACGCTTACAATTTGCCATTCGCCATTCAGGCTGCGCAAYTGTTGGGAAGGGCNATCGGTGCGGGCCTCTTCGCTATTACGCCAGCTGGCGAAAGGGGGATGTGCTGCAAGGCGATTAAGTTGGGTAACGCCAGGGTTTTCCCAGTCACGACGTTGTAAAACGACGGCCAGTGAATTGTAATACGACTCACTATAGGGCGAATTGGGTACCGGGCCCCCCCTCGAGGTCGACGGTATCGATAAGCTTGATATCGAATTCGGCACGAGCGAAGCCAGGGCCTTGCACTTCCTAGGCAAGCGCTCTACCACTGAGCTAAATCCCCAACCCCTTGTTTTATTTTTAAGCAAACGAGATACATAATTTCARCCATGATAATTTAAGATTATCTTGAACTCTTAAGGAAATGTATATACTAAGCTATTATAGTTTTTATTTTCCCTAATTCAGTGGCATAATACCTTACCTTGAGTCGTTTACTACTTTCTTTGGTTTCTAAAAAACTCTACTGCTAAATTACAATGTAAAAAACATAGGGCTCGTATATACTGTAGAGTGCTGTAGATGTCCTCGTACATCAACTATGCAAATAACAGTCTGATCGACACATTTCAGGAKCGATCACTCTTTGGTGTGCTTCTTTAAATACTTTCAGAAGCTTAGGATGTGCAAAGCAGGAAGACTGTGGGTGTAAATGTTTACTTATTTCTTTGAGAGTGTTAGTAACTCTTTTCDAAATTGCTTTTCTCTTCAAAATTATCGTTAACTTAAATGATAATTATCTTTGAGGTTAAACAGAAGCTCATTGACAAACTAAAGTGACTTTTTAGGGCATTCTTTGAGATCATAGTCTTATATCTTGGGGACTAAAATGTCATTAGACCCTAATAGACTAACTTGTATGTTTGTGTGGGGAAACGTTTTCCTCTCTCATTCAAGGTAACTGTTTGCTGCCTGTTGTTACTTGTGTAGCATTCTAGAAAATGGCTAGGTTTTTTATAAGATTTAAGACAATAGAAGTAGTTTTATATTATTATAGTTCTGTTGGAATGTGATCCTGAAATTATTACTGAAAATTAGAATTTTTATTTCGCTAATGACAACCTTGACTCTCAGAGATGCAGTGTAAATTGATACCTCATCTTTCCGAGAGTTCAGAGCACAGGGCGGCAGTATGTGAAGCTGCTTTTGCACTGACGCATTTTGATAAGTTTGGCTACTGTAATGGTAAAAGGCTCCTCAGGCACTGACTGCATTTGGGTTCTTCCGATGGGGGATGATCCGTTCTCGTGGTGCTGCTGGACTTATGCATTTTGGAGGTACTGCATGTATCTTCCACACTGCTTGACATTTTCTCTGATCTGTGTGTTTGCACCAACTCATTAAAAGAAATATGCAGAAATATCTTCTAATTCGTTGATCTTCGCTGTATGACAGTTATAATATTAAACACTTGGGTTGATCCACTCTGTTTACATTTATCTTTCTAAGCGTCAGAAAGGGACTAACTTGAAATTATATCTAGAGGCTTTGTATCATTTCAAAAATTAAATTTCCTTGGATACTTTAGGCAATATCTTAAACAACTTTTTAATAAATTTAAATATTTATATTTACGTAAGCTAAAATATACATGAAATGTGCTTTTTAATAAATTAAATACAGTTTATACTTATTTGCCAATTCACAAATAAAAAAAAAAAAAAAAAAAAAAAA-3′.

[0130] This clone is similar to GLGF-domain protein Homer (accession #U92079).

[0131] Another IEG nucleic acid clone was designated A021. The followingnucleic acid sequence is within the A021 clone: (SEQ ID NO:6)5′-TTTTTTTTTTTTTTTTTTTTAARGGGRCCACCCCACCGSGCTAAAGGCCCAGGGGCCCCCCCCTTGGAGMCCCAGGGGTTTTGGCCCMCCCCCTCACCCAAATGGTCTGCCAATGACCCAGGTACTCACAACATGTTCCAGGAGGAGMCTGGGGCCAGGATTTTGACCAGAGGGTATGGGAAGGGAAAGGGGAGAAGAAATCGACATTTATTTTTATTATTTATTTTAAATGTTTACAWTTTCTTTGTGTTGTTCCAAGCCCTGAATAGAAACAGATAGCATTAAAGGACTCTGTTCCCACCCCTTCTCTGTCTCTCTCTCCCCCACTTGTGCTAACTTAGGATAACACTCTCTATTTCGTTTTGTTTCTAAAGTGATTTGTGGACTTGTGCCGTGTGAACTGCATTAAAAAGGTTCTGTTTTCAAAGATCGATTGTCGTTCCTGTGGGGACAGTGGCTCCTAAGAAATCTGCATTGTAGGAGAAGACAATGAAAGACCTGGCCCTGTCTCTCAAAACTTAACTCTCTGTATGATTTAAAAAAAAATTCCATTTACTTTACTTTGTGGTTACTTGATTTTGAGGAAGAAAATATTCAACTTTGTATAAAGACTAGGTATCAGGGTTTCTTTTGCAGTGGGAGTTGTATATATATCGTATTTTGGTATATCGTAGAAACTCAAGCTTTATGCATCCGTATTTGGGATATGTCAATGACGTGCAGTGAAATTTGCTATTAGACCCTGGAGGCAAACGAGTTGTACAAGGTTTTATGGCTCCATGGGGAATTCTAATTTCCTTTCTGGGGACCTTTTGTCCCGTTTTTACAGTAATGGTGAAATGGTCCTAGGAGGGTCTCTCTAGTCGAATTCTCCAGGCAGGACCACGTGCTCAAAAAATCTTTGTATAGTTTTAAATTTTTGAGGAGTATCTCTGCTCAGAAGCATCTGTGGTGGTGTGTGTTGCGTTGTTCTGTGTACTGTGTGTGACACAAGCCTACAGTATTTGCACTAAGGAAAGCTGTTTAGAGCTTGCTGCTATGGAGGGAAGAACATATTAAAACTTATTTTCCCTCGGGGWTTRTWCWMGTTTTATGTWCTTGTTGTCTTGTTGGCTTTCCTACTTTCCACTGAGTAGCATTTTGTAGAATAAAATGAATTAAGATCAGMWRWRWRMAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAA-3′.

[0132] This clone is similar to fra2.

[0133] Another IEG nucleic acid clone was designated A024. The followingtwo nucleic acid sequences are within the A024 clone: (SEQ ID NO:7)5′-TCAGGCCTNAGCAATCCTCNTTAANTTTGANCCAAGNTTAACTCTTGGGGCGAATTCCTGTGNTTGCTTTCTTTCCCCATANTTCCAGGCCCACAAANGGTTTCTGTGANTCCGAGAATCGGCCCACCATGCAGACCCACNGAGAGGATTCAGAATGTGTGAGAGTGAGTGTGTGAGTGCGCGTGCGTGTGCTTTGTATGTGTGTTTATAGATGTAGGACATTAACTTCCTTCTGACACAGGGAAGATGTGAGAAGGATGGCCTGACATCAGATGACAAGAGGTCTTATAGCACATCTCTGGGCTTTTCCCTACCCAGAGAAGAGCCCCCTTTGATACAAATCAGTTGGATTTTCATATGCTTCAAAGGCTTGATCTGTGAGTCACTCCAGTTTGGGACATAGGTCTGTCTGTGGCTTTGAGAAAAGGTACTTTCAAAAGAGGGCTTTCCAGAGCACAGCTCACAGCCAGCTGTTAGGACCCCACCCTTCTCCTTTATTGTGGAGGTGACTCACAGCAGACTGACAGTGGTCAGACTGAGCTTTCTGCTAAGGTGGTGAGGTAGCCAACACTGGCATGTCTCGGTAGTGGTTTGGGCAAATTTCCGCAGGTCTCTTCCCCCAACCCTGCCTCTGATGAATAAAGACAATGAGTCAGTTCCTTAATTCAGGCTTTTGTGACTAGCTTACTACGGAACCGAAAATGGTCCCCTTTGTACAAGCCGAGCTGTTATGGAATCACGGTGAACCAGACCCAGGTCTGTGGCACCTGTTTGTTTTTTTTTTTTTTTTTTTTTTTTTAGCTCTCATTTCTACGGCATGCTTTCCAAGGAACCAAAGGAGGGTCTCAGAGATGCCCCAAACATCCCAAAGTACACAAAGCTAAGTAATCGATTGCTTACTTATTGCACAGCTAGACACGGATTTTAAGTCTATCTTAAAGCTTTGAAGCAAGCTTAGCTTCTCAAAGGCCTAGCAGAGCCTTGGCACCCCAGGATCCTTTCTGTAGGCTAATTCCTCTTATCCAGCGGCATATGGAGTATCCTTATTGCTAAAGAGGATTCTGGCTCCTTTAAGGAAGTTTGATTTCTGATTCAGAGTCCTTGTTTCCCTGACTTGCTCTGCCAGCCCTGCACCAGCTTTTTCGAAGTGCACTATGCTTGTGTTTAACTTCTCCCAGTTTTATTTGGGCATAAAAGTTGTTGCCTTTATTTGTAAAGCTGTTATAAATATATATTATATAAATATATGACAAAGGAAAATGTTTCAGATGTCTATTTGTATAATTACTTGATCTACACAGTGAGGAAAAAAATGAATGTATTTCTGTTTTTGAAGAGAATAATTTTTTTCTCTAGGGAGAGGAGAGGTTACAGTGTTTATATTTTGAAACCTTCCTGAAGGTGTGAAATTGTAAATATTTTTATCTAAGTAAATGTTAAGTAGTTGTTTTAAAAAGACTTAATAAAATAAGCTTTTTCCTGTGAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAA-3′ and (SEQ ID NO:8)5′-GTGGCCCCTGCTCGCCGCATCATGGAGCGGATCCCCAGCGCGCAACCACCTCCTACCTGCCTGCCCAAAACGCCAGGGCTGGAGCACGGAGACCTGTCAGGGATGGATTTTGCCCACATGTACCAAGTGTACAAGTCCAGGCGGGGAATAAAACGGAGCGAGGACAGCAAGGAAACTTACAAATTGCCGCACCGGTTGATTGAGAAAAAGAGACGTGACCGGATTAACGAGTGCATTGCCCAGCTGAAGGATCTCCTACCCGAACATCTCAAACTTACTACTTTGGGTCACTTGGAGAAAGCAGTGGTTCTCGAGCTGACGCTGAAGCACGTGAAAGCATTGACAAACCTAATTGATCAGCAGCAGCAGAAAATCATGGCCCTGCAGAGCGGTTTACAAGCTGGTGATCTGTCGGGAAGAAATATTGAGGCAGGACAAGAAATGTTCTGCTCCGGTTTCCAGACCTGTGCCCGGGAGGTACTTCAGTACCTGGCCAAGCATGAAAACACTAGGGACCTGAAGTCTTCCCAGCTTGTCACTCATCTCCACCGTGTGGTCTCTGAACTCCTGCAGGGTAGTGCTTCCAGGAAACCAGGACTCAGCTCCCAAACCCGTGGACTTCAAAGAGAAGCCCAGCTTCCTAGCCAAGGGATCAGAAGGCCCTGGGAAAAACTGTGTGCCAGTCATCCAGAGGACTTTTGCTCCCTCGGGCGGGGAGCAGAGTGGTAGTGACACGGACACAGACAGTGGCTACGGAGGCGAATTGGAGAAGGGTGACTTGCGCAGTGAGCAACCCTACTTCAAGAGCGATCACGGACGCAGGTTCACCGTGGGAGAACGCGTCAGCACAATTAAGCAAGAATCTGAAGAGCCCCCCACCAAAAAGAGCCGAATGCAGCTCTCAGATGAGGAAGGCCACTTCGTGGGCAGTGACCTGATGGGTTCCCCATTTCTTGGGCCTCACCCACATCAGCCTCCCTTTTGCCTGCCCTTCTATCTCATCCCACCATCGGCCACTGCCTATCTGCCTATGCTGGAGAAATGCTGGTATCCGACCTCTGTGCCACTGTTATACCCAAGCCTCAACACCTCAGCAGCAGCCCTCTCCAGCTTCATGAACCAGACAAGATCCAACTCCCTTGCTCTGCCCAGAAATCCCTTCTCCCTTGGCACATTCGTCCCTTGACTCTCAAGCCTGCTCAAGCCCTGAAGCAGATCCCTCCTTAAACTTAGAAACAAAGATAAACCTTGAGGGCAATCNCTGCGCCTTGCTTTCCTTCCCACAATTCAAGACACAAAAGGTCTGTACTCAAAACAGAGAGATCAGCCCACCCTGCAGACCCACAGAGAAGATTCAGAGTGTGTGTGAGAGTGAGTGAGTGTGCGTGCGTGCGTGCTTGTATGTATGTTTGTATATGTAGGACAATAAGTTCCTTCTGACACAAGGGAGACACGAGAAGGATAGCCTGACATCAGATGACAGACTGGAGGACTGTAGCACATCTCTGGGCGTTTCCCTACCCAGAGAAGAGCC-3′.

[0134] This clone is similar to a basic helix-loop-helix polypeptide.

[0135] Another IEG nucleic acid clone was designated L003. The followingnucleic acid sequence is within the L003 clone: (SEQ ID NO:9)5′-GCACGAGGGAGTTTATTTCCACGTCTCTTAGGAAAGCCTCGCTGGTTACACATGGCAATGATTGCJGCAGATACACGTCTTAACACCAGAGTACAGTACACACACATTGAGCTGCCCTCGTCTCAAGCAGTTGCAGTTTGTTTAAATGTGAATATCTATGAAACGAGCAAAGCAACTTTCCAGAGTATAGCTTATCACAGAATAGTAACACATGGGCCGCTACTGTATCATACAGAGTACCTCTATAGCTTTTCATCCCCGTGTGAGCATTTCCAAATCACTCAATGAGCACCAAGCACGGACAAGTGACTAAAAAGGCTAGTCCCAATCTCCCCGCAACCCTCGGCGGTAAGGGTAAAGAATTTTGTTTCAAGTAAGTTTTCTCCTCGTCTCTCTCTTCTGAAGACCTGAGCAAAACCAACATTCTAAACCACCCCAAGATATGATACTAGAATTTAAAGGCCCGATGGCTTCAACCCAGAACCTTAACCTACTAGATAAAATCTCTCCGAATCTGACTCACTGATGCTGTTAAGTCCGACAGTACAATCACATAGTACCTCTTTGATACTGTCAAAGTTGGTTTTAAAAATGCCCTAAGAAAACCAAATCATTTTTGGGAGATGTTCTAAGCAAGCTTTCCAACATATAAAGAACAAAACCATGTTACTAAAAACATGGTGCAGGTCCTCACAAAACATTTACTGCTACTACCAGGAAACCAAGCTACTCTTGGTTGTGCTCCTGGTGATAACTGGTGAGCTTTGGACAGCTGCTGGCACATGTCCACTGTGTTCCGTTTTATAATCAAGTGTCAGTTTTCCACTCGACAGAGATTAAAGACAATAGCTTAAAAGTGAAAATGAAATTTCAAGTAGAAGCTACAATTGAATGCTACTTGTTGAGACTTTTAACTTTCACATCCAAATATCAAAAACTTAACTTTGACGACACATGCACACAACACACCATTTGGGAAAGGGTCTTGTTATGCAGTTCAAGCTGGCCTTGAACTCATGATCTCCTGCCTCAGTTTCTTGGGCAGTAGCACTGGACCTTACTGTGGGCAGAAAGTATTGCTCCAATTAGAAAGCATTACTATACACTTCACTTCGTCATGTGCCTAGTGTGGCTCTGAAGGCATAGGAACAATGAAATTAAATTCTTCAGCAGCTGAGGATTCTCTATACTTCAACATTCTGAACTTCAATCATGGCTTCACATTTGAGGCTGAGCTAGATACAAAAATATCAAAACATCCCATAGAATTGTTTATTTCCCTATGTTACTGTTTACCCAAGGAATGTGAAGACTAAAAAAGGACTCATTTGGTTGTTTAATTATGATTAAATTATGTAAATATACAAACATTTAACAAAGCCATCATATTCCAATCTTTTACGAATTCTAACTGCTAGCAGTTGAGCAGCTTTAGATACACTAATAAAATATACAATTTAAAATAGTCGCATTCAATCCTACTAACTTTATAAATAACTTCTTAGGTTAGACTTCTTCCTGCCTAGTTTATAAGACAGTCTAAACCCAAAACTCAACACATATTAAGCTTTTTAAAAACTCCATATAGTTCTAAAGTAACCTCAATGTATTCCCAAGAACCGCCACCATCAATCAGCTCACTCCCTCACACCACTGACTTTAAGACGCTCCTGGGTGGAGAACTGCCAGGCAGAAGCTCTACCTTTCTAGTGTGTGTGGTGGTCTGCTGCTCCTAGTCCAGATCTGGACCACATCAGCACAGCATCAGTGTGACTCAGCACTGAGGCCTTGAGCGCTCTTCCCCCCGATGGCCTGTGTATAGAGGTGTCTAATTCCTTGTGTATAGATGGCCTGTATATAGAGGTGTCTAATTCCTTGGCTCTGTATGTATAGGTAATGTGATACTTTACCATTAAAGCACTATTTTCTCCATTCAAGAATTTAGTGATATAGGAAAATGAGTGGACTTGCGAGACTCAGAAAAACAAAACATAACCTGTCTTGAATTCAAAACAAACCATGGGTGTAGGGGGGAACTGATGAAAGTTTATGGGTTTAACTCTAGGTAATTAACTAAGACAGTCACGAAACACATTATCAAAATCCTTTCAGGCCCAGAGCTTGTACTGTACCCCACTGTGAGACCACATCACAACCCCGGATTGAGCTTTATCCACAACACCTACACCATAGTAACGCAAAGTGCACAATGTACTAAAATAAATTCCTATTAGTTTTATGCAAACTATGGTATAAAATTATCACCTGCCATACATATTTTGCCATGGCACCAACTTCATATAATAAGCCAACGTATAATCAAAGTCCTTACCAGCACCAATCAATGTCCTTGGCACCACTGGACACTCACCGTCAAGCTGTTCATCTAAGAGCCAGTCTGTTCTGACCTGAACAGTTGTGCATTCCACCTTACCACACCCAAGTCTGTGAGCCGGACAAGTGTTTAAATGCAGTTTTACATCTAACGGTGCAGGTTAAGCCGAGCACTTGAAACTGATCACTCATTAATACCTGTCTCCCTCCATACATGTACACCACATGTACACAGCTATGTGCTCTGACTTCAGAATAGCTCTTCCTGTTGGCAAAACACCACAGACATGAAGGGGCCTAGTGTGAAGCGAGCTCACAGAATGTTGGATGGAACTTCGACTATAATGGAAACACCTGCAAAAGCTTTGCTAACCCAGCAAACACTCAACACTTACCAAAGACAACAGGGAAGTTAAAGTTAGCTCGCCAAGAGATGGGCTGGGGAGGTGGGGGTGTAACTCAAAGAAAGCTTTAGCTAACAAAAACGAATGATGGACAACTTCAGAAATTCCCTAAAAACAGAACCTGAAAGTGCAGGTGAGGTTTTGTCCTTCAGTAACAAATGCAGACAGATTCCCAACAGGAATAAAACAGTCTGGGGCTTTGAAACCTGCTAGATGGAAACACGAACTCAAAATGTGGAACCAAGGAAAACCAAATACTTAAATGTGTAAGATAATTTATAATAGTAAAAAGTTGCAAATTGCTGTGACTTGATTTGCCGAAAACATCTGTAAATCCACACTGGCAGTTAGAAGACCAGTTCCCACATTAACTCCTCTCTCAGCAGGTAACCGTTTGTGCGCAGAAGTATCTGAAACATCGCACTACTGCTTATTTTATGGTGTATTGTGCAGAATCTGTACATGCTATTACAGACAATACATATTTGTAAACCTGGTCATGCAAAATCAGTGTGTACAAGGGGATATTGTTAAGCCTTATAAAGTGGTACTTTATTATCTTTGTGACGATGCCAATCTCTCCGAAATATAGCATATCTTAAATGGATATTCTTTATCTGCCAGTTAAAATCATTTTATGTCACTGAAAGAAGAGGTTATACAAGGAAAGAAACATGGTCCTTGTGTTGCAGAATTGATTTTAAATGAGAGAATTTACAAAACCAAGAAATCCATGGTCATAAAGTTTTAACATTTTAATCCTACACATTACAGGGCAAACAGATACTGGACCCTATTTCCACATTCCATAAATCCAAACTTTAGTTCCCATTTCAAACGTTGCCCTAACCACTAAAACCATCAGTGGTCTTACAACCTCTGGATTATGGAAATACAGATTTCTGAAGTAAAAGCTACAAAAAACAACAATGGAAGAAAGCTGAACAAACTTCCCATGAATGAAAATAAAAGTGGAACATCCTGAAGCTCTAGACACTTCTCTCCCGTGTCTATGGTCAACTTGTCGGTTCAGTGCACTGTGCGGTCAAATGTAATGGTCCTCATGTGGAACACACGTCTAACTAGTGTCCATTGATTCCAAGTTAGTGGACGAAGAATCTTTCTGGATACTTCAAAGATGGCTGCCAGCTCCGGGTTGGAGCTGATCTGTGACTGGAACTCACTCATGAGAGGGCTCTTCTCTGCCTCTGGAATGGTGAGCAGTGCAGCTACTGCCCTCATGGCCGAGCGCTTTAACTCGTCCTGCTTTTCAAACTCCTGCTTTACAGAGTTCGCCTTCACCTTAGTTGTACACGTAGCTCGTAGTGGCTCAACAAGCCGGTCCAACCTCTGTAGTACTGCACTTGGACAAAGGGTAGATAGTCTCACCAACATTAAAAATGTTAGCATCTTAATATCATAATGGTCCTTCAAAACCATCTTCCACATGATTTAGAAATTCAAAGATATCCAGTCTGTCAAGACAGCTGTCTAGAAGTGTGTACATACACTCAAAAGCTGCCTTTCTAATGTCCAGGCCGTCATCAACCGTGTGCTTAAACGGGCCCATCTCTACCCTCTCTTATAAAGTTCCTTCCTAACTTTTGTCTCATTGTAAAGATGTGGAAGAACAGAATCCAGAAGGTCCCGTATCAGTGACGGCTTGTTATGGGCTGCAGAATTGAATGTGACCAAGGCCACTCTTCTTACATTCAAATCTGGGTCTTCCAACGTTTTTAGAAAATCACCTATGCAGTTCTTGAGCAGTGGATCTTCTTGACATCTTGAATAAACTGACCTACTACAGCCGGTCCCTCTTTAGGGCATGCTCGAGTAAGGGCAGCTACACATTTGGCAATGGAATAGTAAGACTGCTTATGAGTAAGAGCTGTGCTCTGAGAGTAAACTGGACCCGTTAGCATGCGCAGCAAATCCATGTATCCTAGATTGTTTGTTCCAGTGACAACCAAAGCTLGGAAGAAGTCTAGCATGGCACTAAGAGCTCCTCCCTGCAGCAGAGGTGACCTTACAAGTCCAATCAGTTCATTGAGAATAGATCCGCTTATCTTTGAAAGGGAGGAGGGATATACTTTTGCCAGGGTAGTAAGGAAGCTGATAGCCATCTGGGACACGTGCATATCACTTTCGCTGATAGAGGAGGGAGCTCATCCAGAACTGCATCAATCATGGCGGCCGTCAAACTGTCACTATAGTTTTTAATGAGAATATCTAGGGCAGAGAGGGTCCCCAGTTTCAAAGCTCTCTGATTTTTCCTGAGAAATGAAGCAAGGATAGGGACTCCCTCTCCCAGCACAGGCCTCAGATCTATCTTCAAAGGTGACCCAGCAATCAGGGTCAGTGCTTTCACTGTCGTTAGCCGGGTGATTTCATTCTTGAGTCTCTCCAAGAAAATCTGAAGTGTATTTGATAAGTCAGGGCCCAAATTGTCTCCAAGATTGCAAATAATCTGTCCCATACAGGAAATAGCCCTCTCCTTGACTTCCTGATCAATGTCAGCTGCTTTTAAGCGCTTAATTGTACAAGTGAAGAGATCTTTGATGTAAGGCGTTGCATCGAAGGAGGAGGGTTGGTCCAGAGGACGGATTACTTTGACAAGCTGCTGAGTGACAAGAAGGGCTTCTGATGTGATCTTGTAAAATGGGTCACCAACACAAGCCACCACTGGAGGGACCAAAGCCTGAACATGCGGGTGGAAAACTTGCG-3′.

[0136] This clone is similar to a TATA-binding polypeptide (TTP120).

[0137] Another IEG nucleic acid clone was designated L048. The nucleicacid sequence of the L048 clone is as follows: (SEQ ID NO:10)5′-TCGCCGCCCGAAGTCGCGCAGCTTCCCTGGCGAACGCGGAAGCCCGAAGAGCGCCGTCCTCGGGCCCTGTCGGCGCTCAGGCCCCTTCGCGCGCCTCCTCGCTCGGCCGGGACGTTGCTGTGGAGGCGTGAGGCGCCGGCGGTCGAGCACCTGGAGCGACGGTAGCCCGCGGCCTGCGGTTCTTCTCCTCCCCCGCCGCCCTCCCACCCGAGCTGCGGCGGGGCTCGGCCGCCTCGGTGCTTCTGCACGAACAAAGGAGGCCCCCGCGGCGCCGGCGCAGCTCCATCTGCGGTCCGATCCACCCGGGCCCGCGGCGGCCGCTAGCCAGCCCTTCCCGGAGGCCTCAGCCCGGCCCACCGCCCGGCGTCGCGCGCCAGCTCGCTAGTGCATCCGGGCCCCGCAGGCACAAAAATATGGCTCAGGAGACTAACCAGACCCCAGGGCCCATGCTGTGTAGTACTGGATGTGGCTTTTATGGGAATCCTAGGACAAATGGAATGTGTTCTGTTTGCTACAAAGAACATCTTCAGAGACAGCAGAATAGTGGCAGAATGAGCCCAATGGGGACAGCTAGTGGTTCCAACAGTCCTACCTCAGACTCTGCGTCTGTACAAAGAGCAGATGCTACTTTAAACAACTGTGAAGGTGCTGCTGGCAGCACATCTGAAAAATCAAGAAATGTGCCTGTGGCTGCCTTGCCTGTAACTCAACAAATGACAGAAATGAGCATTTCAAGAGAGGACAAAATAACCTCCCCGAAAACAGAGGTGTCAGAGCCAGTTGTCACTCAGCCCAGTCCATCAGTTTCTCAGCCCAGTTCTTCTCAGTGAAGAAAAAGCTCCTGAGTTGCCCAAACCAAAGAAGAACAGATGTTTTATGTGTAGAAAGAAAGTTGGCCTTACAGGGTTTGACTGCCGATGTGGAAATTTGTTTTGTGGACTTCACCGTTACTCTGACAAGCACAACTGTCCTTATGATTACAAAGCAGAAGCTGCAGCAAAAATCAGAAAAGAAAATCCAGTTGTTGTGGCTGAAAAAATCCAGAGAATATAAAATTACTACATGTGAAGAGACTGAAACTTTGTTTTTATTTTAATATATCGTAGGAAAACATTAAAGAGCAGATGCATGGCCATTTTCCTTTGATGTTCTCCAGAGTTTTGCTTTATACTTGTCTGTCATATAATTGATATTTTAGGATGTTTGGGTGTTTGTTACAGGCAGAATTGGATAGATACAGCCCAACAAATGTATAGCCCTCCCCTCAGTAAAATTGGACAAAAATATGCACAGCAAATTGAAATACACATATACTAGGAACAAAATTTAGTTCCATGTGCCAAACTGAATGAAATCTCTGCATGTTTGCAGCATATCTGCCTTTTGGGAATGTAATCAAGGTATAATCTTTGGCTAGTGTTATGTGCCTGTACTTTAAAAAAATGGTACACCAGAAAAGGACTGGCAGTCTACTACCATAGTCAAACTTCACCTTAATTTCGACATGCTTTTGGAAGCAGGAAGAAAGCTACAAAACCAGTATTTGGTGCCATGTGTGAGCCTGGTTAAATTGGTCTTCTAAAAGCTGTCAATTAGGACATTCTGCGAAAGGTAACATCACAACTGGTTCTGAGTAAAACCATCAAGTCAACAGCAGGGTGCCTGAGATAATCTTTGAAGCTTATTGTGCTGGCCTGCACCAGAAGATATCTGCATTCTCATTACTAAAATTGTAGCACAGAACTGCACTAGGATTTGTTTACAAGAAGAAATTAAAACTCTACGTTTGGTTTTCACATATAGCAGCTCTGTTAAATAACATGCATCTGAATTTTAAGTTGCAAAGGTATCTGAGCAGTTAGTTTTTCATGTGCATCTTTTGTTGAATGTTTTGGTTCAAGAAAGAATGTTTAAAGCTTTTTAAAGACTTCAGTTCTTAATGTAACTGTACCCTTCTGCATGGAAAATCATAACCAACATGGCTGCAGTAGACTTCTTTAGTGGTATCCAGCACCACTTGCAGAGGGCTGCTTTATCATATTGTATTTGGGTGTAGGACTCTAGTGTTCTTGGGTGTATTGCATGGGCTGCATTATCTACAGCATTGTACAATAACAACTAGAAAAGGCAGTATACTTCACTGATGCTTGTCTGGTAATATCACTTCTGTGTTATAATGGAAGGTTTTTTGTGATGTATGAAACTTGTGTTTTTTATATATAAATGAGTATAGTTAGATTAGTGTTGTGGTAATGCCTGTTTTCATCTGTAAATAGTTAAGTATGTACACAAGGCACTACTTCTGATTTATTGCAGTGTTCAGTCCTAGTTTTTCTTTATTAAAACATTCAGTTTTGCTTCAATTTTATGTACTTTAGTTCTAAGTTAGATTTGCAGATGTGTACAGATAGTTCATATTTATGTATTGCACATAATCATGCTATTCAGCATTGATGCTATATTGTATTATGTAAATAATAAAAGCAGTGTACAGAGGGAAAAAAAAACTCGTGC-3′.

[0138] In addition, the L048 clone contains an open reading frame (ORF)from basepair 414 through basepair 1055. This ORF encodes a polypeptideof 214 amino acid residues. The amino acid sequence of the L048polypeptide is as follows: (SEQ ID NO:11)MAQETNQTPGPMLCSTGCGFYGNPRTNGMCSVCYKEHLQRQQNSGRMSPMGTASGSNSPTSDSASVQRADATLNNCEGAAGSTSEKSRNVPVAALPVTQQMTEMSTSREDKTTSPKTEVSEPVVTQPSPSVSQPSSSQSEEKAPELPKYKAKNRCFMCRKKVGLTGFDCRCGNLFCGLHRYSDKTTNCPYDYKAEAAAKT RKENPVVVAEKIQRI.

[0139] In addition, the L048 polypeptide was found to be cysteine rich,having a motif with distant homology to that of polypeptides withZn⁺⁺-fingers.

[0140] Northern blot analysis using a sequence from the L048 clonerevealed the presence of a 2.5 kb mRNA transcript. In addition, thisanalysis revealed that the expression of the L048 mRNA was stronglyupregulated in response to the multiple MECS treatment.

[0141] Another IEG nucleic acid clone was designated L064. The followingnucleic acid sequence is within the L064 clone: (SEQ ID NO:12)5′-ATTCCAAAAATGCATAGATTACAAAGAAACACCAGACAAGCTCAAACTCAAGGATATTCTACAAATAAACCAGTACCTTCAAAATGCCATGCTACCAGGTACAGACAGGCGAGANACTGTTCCACACTGAGGAACTAACAAAGTATCCATGAAGTCCATAATTGTGGGTCAAATCCAGGACCTGCAAAGGGGATTTGGGGATAATTTTCAAAATTTGACTAAGGTCTGCAGAGTAGAGAGACGAGGTCAATGCCAATGTCCTGATTTTGACAGTAAGTATTTAAATATGCAGGAGAACAACCTAACCAAGAGGCTGCCAACACACTTCCTGGCTGTGGCACAACTAGATTTAAAACCAGCAATTTGTTGGTTCTTGTTCTCAAATATCAGTTACCTGCAAGCACTCCATCGTGAAAGGATTGAGAGCATGAGGTGATGTGTTGATGGTGAAAATGAGAACTGACTGAGCACAGGAAAGAGTGGCATGATGGGCAGGGAAAGGGGAGACAAAGGTCACAAGAGCATGCAACACTCAGTGAACTACAGGACACTCCAAAAGGCACTCTGCTGTCTAGCTTGGATCTGGAGGAGGATCAGNTATTAATAAGGGCCCTGGAAGGGNCAAAGCTAGCCTCCCAGCTGCTGGC TTCCCATCTGCT-3′.

[0142] Another IEG nucleic acid clone was designated L067. The followingnucleic acid sequence is within the L067 clone: (SEQ ID NO:13)5′-GCCACCACCATTGTTAATGGAGGGAGGCTCTCCCTTGTTATTTCTCAGAAGACTGAATGTCTGTACCAAAAGGCTCATGGCTTTCTCTGGGCCTTTCCATTTAAGGTTATAGTTTTTTATGTAGTGTTACTAAAATCTAGGCTTGTTACTAAAGTGGGCTTTGTAGTTATTGGTATCGGTGGATTTTTATGTTACTTGGAGTCCAGAACAGGGAGAGCTCACCACAAACCTCTCCTTTCCCTGGACCAAACACCCTCTCTGTCCTGTGAACTCACCTTTTCTTCTCTGTGGGTCACTCCCATTACCACACTGGTGAGCGAGCCAAATGGATGAGAGACACAAAGACCGTAGTTCTTGAGAGACATTATTTTTTTCAACTTTGTTTTTTAAGAGATTTTATGTGTTGATTTGTTTTGGTTTGGTTTAAAGGGATTCATAGCTAACTTGGATTTTTGTTACCTCAGCTCTGGGAGAGGATTTTTGCTGAATGACTATTAATTACCTGAGCATTGTTGCTCTGAGGTCATGGCATGCTAGCCTATGTCTGTTACAGTCTCAGGCTGCCCTTGTTTCCTCGTTCCTGTGCTATTGTGCTACACGCTCAAGGGGCCTTGACTCTGCTTACACACATTAGGGGCAGTGTGAGTAAATGTGCAGTGTCCACACTTGAGGACATGAATGTCTGCACTGTCACTTTGCTCTGGGTGTGAAGTCCCTGGTCCCCTTGCTCCTGTAGCTTTCTTTTGATCGACTACTGGAACTCAACCCTGTGTACAAGAGCAGCACTGCCTCTGGTGGGTGGTGTTTGCAGCCAGGATTAGATGCCAGTCCTCGGGTTCCCTGGCCTTGTTGGAAAGGTGTGCTTCCTTGAGGTCTGAGAATGGAAGGCTCTGCCTCACTCTAGCTAGGAGGCGCAATGGGAAAGTATGAGTTCAGGGCGTCAGGGCAGTGGCTCCTGAAGAGCCAGCTGTGGACAGAGGGAGTGAGGCTTTATTTAAAGTGACAGGAAGAAACATGGCGTTTTGGTATATTGGGAGCAATGCCAAGATTCCCTCCTGCCCTACATAGGTCACAGACACCTTCCCAACCATCCCCTCCTCCACTTCCATAAATGAAGACAGCCCTGATGACCCTCACCCCTTTTGCATAGGTCACTGGATCCCACTGTCCTTCCTCGGTGCTTACACACTTTACAGACCCTTTAGGCGAGCCCTTGCATAGAGCGTTATCTCAGTGCTCCATTCCAGTCCTGACTCCCTGTGGCCATTGAGACTTTGGATTTAAGAACTCACATTGCTAGGGAGAGGGGCTTTGCTGGGAAAGGTGACTCCTCTGTAACCTAGCCTCTTGTGCTCCTCCATGACAGAAATGCTGGGTGGAGTTTTACATTTGCCAATGGCCAGCTTGTGAATATCTTCATATACACTTTCTATTCATGTTACTGTAGTTTCTGTTTTGAAATAAAACTTCTGAATGT-3′.

[0143] This clone is similar to a glucose transporter type IIIpolypeptide.

[0144] Another IEG nucleic acid clone was designated L076. The followingtwo nucleic acid sequences are within the L076 clone:5′-CATATAAATGTACTTTATTGTTTTAAACAGAACG (SEQ ID NO:14)AAAGAAGAGGCAGAAAACATTTGCATGTAAGTCCTAGCTTATAAATGTAGTTTTTAGTGGTGGCATCTCTAACACGTCGTTCAGGGACTGTTTCCTTTTGCCTCCTTGTACTGTGAGCACTGACACTTGAGAAAAGCACATCTGGCGGACATATGTCTCCAGAACTGGAAGAACTTGGAGAGCAAACATTTTTCTTAATTCCTCTAAGTAATCTTTAGTAAAACAAAAGATGATCTITGGCATAGATTCATACTTTAAAGGCATTGATATGCATTTATATCAGGTAAGCAACTATACAGATCTGCTGAGAGCTTTCAAAAGAATCTGTTATCAGCTGAAAGGAAATAGGGGAAGCCTGAGTATTCAGGGTCAACTTAAGATTTGCAAGTTCAGTGTTGGGGTCAACATACTAGATGTGGGAAGAACATCCAGGCAAGGTCTTAGTCCTGTATTCACCTGGTTCTTGATTTCTGGAAGAAGCATCCATGCGCTAGGAAATGCTTATACAGCCGAGGTAAATGCAAAAATGAGTAAAGTCACTTTTTCACTAACTTTGCCCAATAGGRAACATGCCTTCTGATAAGTAGATACCATACTCTTTATTCTTGAATACTTTATATTGAGAGAAGGTTGTAGTTGGTTAAAAGCAACTGGGAACTATAACTTCCTACTGATTTTTCCCTAGCAGCACCAGAATTATATTCTGCAAATGCTATTCTCCCTTACATAGGAAATATCCTTCAGACAAAATTGCCTTTCCATTCAGTCTCTTAAGAGYTTAATTTTGAATGGACTTTTCAAAGTTACAAGCAAAGTCAAGTGTGGTGGTAGGAGCTAAGAGGCTGACACAAGTAGATGACTTTGAATCCAGAAGTTCAAGACTAGCCTGGACAACATAGAGAGACCCAGTCTCA AAATT-3′ and5′-GGCGGGGATCTCTCGGCTGGTAAGAAGGGG (SEQ ID NO:15)CAGTGGTACCANGCGGGCACTTATTCAGTGTGCCAAGGATATCGCCAAGGCCTCTGATGAGGTGACGAGGTTGGCCAAGGAGGTJGCCAAGCAGTGCACAGATAANGCGGNTTAGAACCAATCTCTTACAGGTCTGTGAGCGAATCCCAACTATAAGCACCCAGCTCAAAATCCTGTCCACAGTGAAGGCCACCATGCTGGGCCGGACCAACATCAGTGACGAGGAGTCTGAGCAGGCCACAGAGATGCTGGTTCATAATGCCCAGAACCTCATGCAGTCTGTGNAAGAGACTGTGCGAGAGGCCGAAGCTGCTTCAATCAAGATTCGANCAGACGCCGGATTTACTCTGCGCTGGGTCAGAAAGACTCCCTGGTACCAGTAGGCACCTGGTCAGACCTGGCTGGTACACAGACCTCTGCTAATGANGANGTGACCATCTTGAGCTTCAGAAGCCATTCAGAGTTGCCAAGGGGTGGNAAATCAATCCCTGGTTTCACACACCAAGAAAGGGAATGGGGCCTCCTTCACATTAGAATAAACATTTATACTCTTGTCATGGGACACTTTGAAAGTGTCTCTCCTACAAAACCCCTGGTACCTTTCAGGNTTACTCCNGGTNGCAANNTCCTCCCCCAAGGGGAATTTTTTACCAATAAAAGGCTCAAGGAATTAANGGCGNTTGAAAACCAACNTNATCCAANGGGAAANGCCCCCNTGGCCTTCTGGCCCCCTTGGGGGNACAATTTTTCNTCCCNCTGGGTGTTTTAAATGGGGTTTCAACCTTGGGGCTGGNCCTTTTTCCNCCCCCCCTTTTAAGGGGCTTCCTCCGAAGGAACCTNAGAAAACTTNAAGGGCCAAAGNTCCANTTTACNAATAACTGGG-3′

[0145] This clone is similar to vinculin.

[0146] Another IEG nucleic acid clone was designated L082. The followingtwo nucleic acid sequences are within the L082 clone:5′-TTTTTTTTTTTTTTTTTTCCTCCCTTAAAAGAT (SEQ ID NO:16)AAACTAATAAACTCTTCAATGGTCTTTTCAGTATAGTTCTTATGTAGTTTAACATAGCTTATAAATTGAGTTTAACAATAAACTCAAGAAGATAATTTTATAAACCCTGTTTTCCAATCTGTCATTTACTTAAATTATTTTGGTTGTTTTCCCTTTTTTTCCTTCTTTTCTCACCCCCTCCCTCTCCATGAAGATTTCAGGTGCTTAACATATCATTTTTTTCCCTGCTGGAATTTTAGCATTGATATGAACCATGGACAAGTATATTCTGCTGCCACAAAGACTGTAAAGTGCTTCATTTCAACAGCTGAGGCAAGCCAAGTGATCATTAATAAAGCTTTTCTTGCTTCCTTCAGTGGTGTTGGTAGTAAAATGGTAGGTAAAAGTTAGGCTGCAAGTTCAATAAATGAGATTTACCTATCATTCCACCCTTGTGTATTCATTCACCTATCCTGGTTCAAGCAGTTTGAGTCAACTAGGCATTTAAAGGCATTGTGTTTATTACTTTATGGTTTCCAACTTTACATACTTGTCAGGGATGAAGTCTGATAGGTTAAGGACAGTAGAAATTTCTGTGCAACAAGCAGCAAC-3′ and 5′-TTTTTTTTTTTTTTTGGTTACAAAAGT (SEQ ID NO:17)ATTTATTTTATAAAACTTGTATTTAAAATAGAGCTTATCTGTCTACTCACAAATCCTAATTTAAAACATAACACATTATCCTTAGCTAATCTGATGTTAACCTTTACAATCAACACTCATTTTTGTAATTTTATTAAGAACCTGTACTAAATGAAGTTTTTAATCAGAAAACATTCCCTTTTATCTTAAAAGTGCTTCTTAAATGAAGGCACCAACAAGAACTACTTTCAGATGGTACAGAATTTCTTATTTCTTGAAGACTCTGTGGTTGACCACTTCTTCATTAGTTACCTGCAGCAAGACACCTTCCTGCCAAAGGAAAAAAAAAAGTATCTGAAGAAGTTTATCATGTTTGTCCAAAGAACCTAAGTAACTTCAGTGGTGGTTTTAGGATTAAAGCAGACTCACTGATGTGTATACGCCCTGAATATCACATTTCTGGAAAGGCAGTAAAGCCTAGAAATCAGAAGGCGGGCGGTTTTAAAGAAATTTCAATAGCCAACCTACAACANTTTAGGGCAAAGATAATGGGCAAAAANTNC-3′

[0147] This clone is potentially similar to a nRNP polypeptide A2/B1.

[0148] Another IEG nucleic acid clone was designated L094. The followingtwo nucleic acid sequences are within the L094 clone:5′-ACGATATMTATWGARRTWTAWCTSTTHAC (SEQ ID NO:18)TGAATMWHATGCACAAATATTAACTAGTRRTTTATTAAACAGATATSATTTAGAACAAGACTTAAWKAAATACAAATCCTTAGGTACGRTTTAATATCATGTTCADGATGTTTGAAGAGTTTAAAAAGAATCACTGATTAAGKKAAGCATCCBCACTTTTCTTTGAGAABCCAAACCTTTTAGGNAAADACCCCATTCCAAATTTTGTCCCCHATTTCAGRCCKKCAGAAAGTCTCTAACATSAAGAGTCCTCAACGGGGNGTAACTCAVAWCTCCTATCAAGTGCAGTAACCTAGCTCTCCCGGDGGCCATGGCGT-3′ and 5′-AAACT (SEQ ID NO:19)AAACAGTGTTTTGTTAATTCTTCTGCATTCGGACTATTGCAGGCATTTAGAGCATCCAGAGCTACGAAGGGCTGGCTGCAGCAGCACCGCCCTTTGTAAGCCAGCAGACCAGCCTTAACTGTGGGCTTGACTCCTGTGAGCTGGCCTCAGTGTGACTCAGAAATGTTTGATTAGCAGATGAGAGAGCGAGGACACACCACGAGGGCTGCGTTCTCTTCCTCCAGCGCTGTGCAGGACAGTTTCTTCTCACCCTAGCCTTTTTAAATGCACCAGAAGTACAGACAGTTGCACTACACAAACCCTTTGAACACTTGTAGAAATCAGTCCACCGTAGATTAGACAGAATCACCTTCCAATCCTTTGACTTCTTTTCCTTTCATTTGAACAATTGTATAATAATTGATTATTGTCAAATTTTTTGTCTGTGGTAGTATCGCTTTAATTTATCTTAGTACATCAACGTTTTGATTTAAAAAAGAATTAAAACAACAAAAAAAGTCACTTAGAAGCCATGAACTTTTTTTTTTNGATNGGGAAATTTTCTTGTTTNGAAAATTATCATTGGGGTTCCTCCGGAAANCTTGTAAGATTGGNTTATAAGGTACCTGGGANGTTTTCANAACNGGTGGNTATACCCTTTTTTAAGGGAAATTAATGATTTNGAGTTTTTGGGCCAACTNCGGGANTGGCAGGGAAACCANNCNGGGGNGGGGTTTTAAATTNTGTGAGGGTTTTTTTGGGCCTNAATTTTTTGCATAATTTTCACCTNGNAACCTTTNAANNCTNGGAAAAAAAAAAAAACNT-3′.

[0149] Northern blot analysis using a sequence from the L094 clonerevealed that the expression of the L094 mRNA was upregulated inresponse to the multiple MECS treatment. Specifically, L094 mRNAexpression was induced 7.3 fold by the multiple MECS treatment asdetermined from Northern blot data using total RNA from rat hippocampus(Table I). In addition, developmental studies revealed that thetranscriptional expression level of L094 was upregulated between day E15and E18, and downregulated at day 0. The expression then increases againduring post natal development.

[0150] Another IEG nucleic acid clone was designated L097. The 5′-end ofthe clone obtained from the first library screen was used to design anantisense primer. Using PCR, L097 DNA was amplified and inserted intothe pCR2.1 vector. The L097 clone is about 4.4 kb in length. Sequenceanalysis of the first 4060 bases from the 3′-end revealed the presenceof a coding region of at least 2351 bp. In addition, RT-PCR analysisrevealed that the L097 clone was missing an adenosine at position 1166from the 5′-end. The lack of this base results in a frame shift in thecoding sequence. Further, the sequence at position 1358 was ambiguous.However, any base substitution at this particular position will notalter the encoded amino acid residue. Specifically, a serine residuewill be encoded by the codon containing nucleic acid position 1358regardless of the base at position 1358. The following nucleic acidsequence is within the L097 clone:5′-TGCAGCCGCCCTTGGAACTGCATGTCAGGAAGCATCCCTTTGTGTA (SEQ ID NO:20)TGTCTGTGCTATATGTCTCAAGAAATTTGTCAGCTCAATCAGGCTGCGCTCCCATATCCGAGAGGTGCATGGGGCGGCCCAGGAGACCTTGGTTTTTACTAGCTCCATCAACCAGAGTTTCTGCCTCCTGGAGCCTGGTGGGGATATCCAGCAGGAAGCCTTGGGAAACCAGCTATCACTGACAGCTGAGGAATTTGTGTGTCCAGAAATTGATGTACGTAAGGGGGAGGTTTGTCCTGGGGAAGCTCAGCCTGAGGTGGGGCTGAGGGAGTTGGAGGCCCCTGGAGAAGCATGTGCCCCAGCCGTGCCCTTGGCCAACCCCCAGAGTGTCAGTGTTTCCCTGTCCCCCTGCAAACTGGAAACCACTGTGGTCAATTCCGACCTCAACTCTCTTGGAGTGGTTTCAGATGATTTTTTACTGAAAACTGATACCTCTTCTGCTGAGCCTCATGCTGCTGCTGAGCTAACCTCAGACACACAGCATCGAGGCTCAGCCCAGACTCAGGGTGAAGAAGTCACACTGCTGCTGGCCAAGGCCAAAAGTACTGGACCAGACTCAGAGAGTCCTCCAAGTGGAGGGCAGAATGTGGGTGCTCTGCCAGCCAGTGAATCTGACTCTAACAGGTGTCTCAGGGCAAACCCAGCAGAGACCTCAGACCTCCTTCCTACAGTGGCTGATGGAGGAGACCTCGGTGTGTGCCAGCCTGACTCTTGCACGTCGTCCTCTGAGCACCACCCTGGCAGCACAGCATTCATGAAGGTCCTAGACAGTCTCCAGAAGAAGCAGATGAACACCAGTCTTTGCGAGCGGATCCGGAAGGTTTATGGAGACCTGGAGTGTGAATACTGTGGCAAACTTTTTTGGTACCAAGTGCATTTTGACATGCATGTCCGCACCCACACCCGGGAACATCTGTATTATTGCTCCCAGTGTCACTACTCTTCCATCACCAAAAACTGCCTTAAACGCCATGTAATTCAGAAACACAGTAACATCTTGCTGAAGTGTCCCACTGACGGCTGTGACTACTCGACTCCAGATAAATATAAGCTACAGGCCCACCTTAAAGTTCACACAGAGCTGGACAAAAGGAGTTATTCTTGTCCTGTATGTGAAAAATCTTTTTCAGAAGACCGATTGATAAAGTCACATATCAAGACTAATCATCCAGAGGTCTCCATGAATACCATTTCTGAGGTTCTTGGGAGAAGAGTCCAGCTCAAAGGGCTAATTGGAAAGCGAGCCATGAAGTGTCCGTATTGCGATTTCTATTTCATGAAGAATGGCTCAGACCTTCAGCGGCACATCTCNGCTCACGAGGGTGTGAAGCCCTTCAAATGTTCTTTGTGTGAGTATGCAACTCGTAGCAAGAGCAACCTCAAAGCTCATATGAATCGTCACAGCACTGAGAAGACTCACCTCTGTGACATGTGTGGCAAGAAATTCAAATCCAAAGGGACATTAAAGAGTCATAAGCTCCTTTCACACATCTGATGGGAAGCAATTCAAGTGCACGGTGTGTGACTACACAGCTGCCCAGAAACCACAGCTGCTGCGACACATGGAGCAGGATGCCTCCTTTCAAGCCTTTCCGCTGCGCTCACTGTCATTATTCATGTAACATCTCTGGATCTCTGAAACGGCACTACAACAGGAAGCACCCCAACGAGGAGTATGCCAACGTGGGCAGCGGGGAGCTTGCAGCTGAAGCCCTCATCCAACAAGGTGGTCTGAAGTGTCCTGTTTGCAGCTTTGTGTATGGAACCAAATGGGAGTTCAACAGACACTTGAAGAACAAGCATGGCTTGAAGCCAGCGACAGAGACTCCCGAGGAGCCCTCCACCCAGTATCTCTACATCACCGAGGCTGAAGATGTTCAGGGGACACAAGCAGCTGTAGCTGCACTTCAGGACCTGCGATATACCTCCGAGAGTGGTGATCGACTTGACCCCACAGCTGTGAATATCCTGCAGCAGATCATTGAACTGGGTTCAGAGACTCACGATGCTGCTGCCGTGGCCTCCGTGGTTGCCATGGCGCCTGGGACAGTGACTGTTGTAAAGCAGGTCACCGATGAGGAACCCAATTCCAACCATACAGTCATGATCCAGGAGACTCTGCAGCAGGCCTCTGTGGAGTTGGCCGAGCAGCACCATCTGGTGGTGTCCTCTGATGACGTGGAGGGCATTGAGACAGTGACAGTGTACACACAGGGTGGGGAGGCCTCAGAGTTCATCGTGTACGTGCAAGAGGCTGTCCAGCCCATGGAGGAGCAGGTCGGGGAGCAGCCAGCCACAGAACTCTAGAGAATCCCTGCCTCCTTTGGCAGCCAGCCTTTGTGGGCCTGAAGACCTCCTAACCCACCAGGTCCATCCCTGGCTCTTCTTGCCCACTGGCCCCAGATAAATTTCTCCATAACTGTCCTCTGTGTGGTCAAAGCCAGGAGAGTATCATGAAGAGAGAGAGAGAGAGAGACTAGTCTCCGAGTTTTTTTTTTT-3′

[0151] In addition, the following amino acid sequence is within the L097polypeptide: QPPLELHVRKHPFVYVCAICLKKFVSSLRLRSHIREVHGAAQETLV (SEQ IDNO:21) FTSSINQSFCLLEPGGDIQQEALGNQLSLTAEEFVCPEIDVRKGEVCPGEAQPEVGLRELEAPGEACAPAVPLANPQSVSVSLSPCKLETTVVNSDLNSLGVVSDDFLLKTDTSSAEPHAAAELTSDTQHRGSAQTQGEEVTLLLAKAKSTGPDSESPPSGGQNVGALPASESDSNRCLRANPAETSDLLPTVADGGDLGVCQPDSCTSSSEHHPGSTAFMKVLDSLQKKQMNTSLCERIRKVTGDLECETCGKLFWTQVHFDMHVRTHTREHLTTCSQCHTSSITKNCLKRHVIQKHSNILLKCPTDGCDTSTPDKTKLQAHLKVHTELDKRSTSCPVCEKSFSEDRLIKSHIKTNHPEVSMNTISEVLGRRVQLKGLLGKRAMKCPTCDFTTMKNGSDLQRHISAHEGVKPFKCSLCETATRSKSNLKAHMNRHSTEKTHLCDMCGKKFKSKGTLKSHKLLHTSDGKQFKCTVCDTTAAQKPQLLRHMEQDASFKPFRCAHCHTSCNISGSLKRHTNRKHPNEEYANVGSGELAAEALIQQGGLKCPVCSFVYGTKWEFNRHLKNKHGLKPATETPEEPSTQYLYITEAEDVQGTQAAVAALQDLRTTSESGDRLDPTAVNILQQIIELGSETHDAAAVASVVAMAPGTVTVVKQVTDEEPNSNHTVMIQETLQQASVELAEQHHLVVSSDDVEGIETVTVTTQGGEASEFIVYVQEAVQPMEEQVGEQPAT EL

[0152] Using tblast2× algorithms, nine Zn⁺⁺-fingers were identified byhomology to motifs of Zn⁺⁺-finger containing polypeptides (accession #PIR2:A32368, S03677, A29634, S06571, and A60392). The presence of themultiple Zn⁺⁺-finger domains suggests that the L097 clone is atranscription factor, however, the size of the encoded polypeptide is inexcess of 700 amino acids.

[0153] Northern blot analysis using a sequence from the L097 cloneindicated that the L097 mRNA transcript is rather rare. In addition,this analysis revealed that the expression of the L097 mRNA was veryweakly upregulated in response to the multiple MECS treatment.

[0154] Another IEG nucleic acid clone was designated L099. The followingfour nucleic acid sequences are within the L099 clone:5′-TGGATCTACTTGTTAATGGTTTCATGGAAGC (SEQ ID NO:22)AATCAGCAATATGTGATATGAACTGCTGCATTACTTTATTATACTCGTGGAACTGAGATATTTARMSRSMGCTTWWTTTTTTTTTTTTTTTAGTGTAAAATACTTAAGCGTTTCCACTATIGGAAGAAAAGCATATATGGGTATTFIGTATTGTAACTTGTTTTAAAAGGACAGTCTTTTTTAATCTTCCCACTTAAATGCTTTTTAAAATATGTAATACAATTTGAAGCTTGTTTAAAAATAGAATTAAATGTCTTAWATAGKGCTACKGTTTTGGAATTAGAAAGTGATCAAATACAAAACATTTTAAAATTAAGCCCAGAAAACAAAATAGTGTTTAAAGTTAGTTTAGTATAAAAGAAATTTATAAGATTTTTTTCTTCAATATAAGATACCTCACTTGAAAATAAAGAAAGCACAGCACATTAAAGTAATTCTCATGAGAACACCCCATTTAGAATAATTGCTAAATCTAGGACACCTTTTGAGTTGTGAGTTTGTGATACATGTAGTCACCATTAGCTTTTCTGCTGGAAGGACTTCCCGTAGTAATTTTAAGGCAGTGTAATAGTTCAATTACCCCACAGTTTCTAACCTGGGAAGGCAGTATGTGAATGGTCCCTTCTGCAACTACGGAAACACATTAGCTACATTGAGCATAACTCGATTGATAATTTTGCCAGTGCATATAGTTTTTATGATTAAAATTGCTGTGGTTGGTTGCATTACACGACACACAAAACTGTCCTCTACCTCACATGAAATAAATATTTTATATGGTTTTACTAAAAAAATGACTCATCTATCTGGTTACTTAGTTTACAAATTTTGGATTATATTTATTGAAACATGACATACTGTGCTCTTAGCTTATACCTCAATCGTATTTTGTGCTGTTTGCCATTTTCATGCCTTGTATATAACTTGTATAGATTGGATGATATTCCCAATAAACACTTTTAATKCCAAWRAAAAAAAAAAAA AAAAA-3′ ;5′-TAATGTTTATGATACAAAGCTACTCACTCTG (SEQ ID NO:23)GAGCCTTCTCATTACAGAATCTCTTGACTTTTATACACCCAGCCTGTTGTTACTTTGTTCAGGTTGCAGAATGAGTTTCCTCTGGTTTCCTCCTAGAGGAGTTTTCCTGATGAAATGCTAGTAGCACCTCCCCGACATACAGCGGGTGGGTGGGGCACACTTTGCTGTGCTCTGATGGTACACACAAGAAGCAGTTGTAATTTGTCTTTCTGTTTTTAAGAGTGACCATAGCTAGATATGTGTGTGTGACTTCAGAAAATTAAAATGCTTTCCGAACTTTTCCTGTTAATAGAGGTGTGAAGTACTCATTCATGTGCATGAGGAAAGTGGATTTCCACGGACGCACACCGCTTTCCTATGTAACTCACAATGCTCTGTACAGTTTTTATATGTAGTCTTACAAAGGTCTTATGAAATTTATATAATGGATTTTTTCTTTTAAATTTATAAAATACTAAATATCTTAAAGATTGTTTTGGACTTTTGTATGTTTAAATGTTATCTTAAAACTTGCACAAATGGACCATGATGACTCTTTGATCTTAAAATCAGGAATTTACAGTCAGCTAAGAAAAATGTGGATAGGTTAATAATCCACAGTGGGAGTATCTGCTAGGAGCAGGAATTGTAGATGACATGAATTCCGTGATTTTGAGGAAGGGCAGCCTCTGCACTTTTCTTTGTTTTTGTTTTGCACATGAAGTCTGACATTTTTACCATCGAATTTCACATTACTAGATGGTTGGCTTGGGATTTACCTAGGGGAAATTCTTAGCAACTTTGTACTTTGTTGTTTTTGTTCTGTTTGGTCTCCAGCTTGCAGAGACCCTCTTGCCTCTGTCTCCCAAGTGTTTGGGTTGGCAGGATGAGCCCCACCACCGCTGGCCCTGTGCAGTTCTTTTGGGATGTCCCTGAAAGCAGCTGTGGCATTATCTTCTGTTTCATGTGTCCCGAGCTGTCTCATGGTACTACATGCAGTGACCTGAGATCTGCGTTAAGGAATAACTTAGGAGAAAACGGCTGTCACTGTCCTCCCCGCTGTGAGACACCAGAGTTATCACACCTGTTATGGTCATACTTTGTGTTATGATACTGATGTCTAAGGCAATTTTTCTACTTTCCAAAAGGGAGTTTGTTTCCTAAATATATTGTGACCTAAATGTGGTTTTATTCTGCTATGTTCTATAATTTATGTATTGACTTTTGTAACCTCCTTGGGAGAAACATGTTTAAGTGGCACAGGGACCATATATGTCATTTTATTTAGCTCTGGAGAAGGAAACCACAGGCGTTTTTGTAAAATAGCATTAGCTTAGATGTCAGTTCATTGTGCTTGGCTGTGTGGGAGGCAGACTCAAGGACTTGCACCATTTATTTTTCTGACAGAAGTGTTCTGCTTATGTGCTGCTTAGTAAGTGTGATTTTTCTAGTCTTGATGAAACTTGCCTCGTGACATTGTTGAGCGTAGTCTTCACTTTCCAGAAGATGAAATGATGTGCCATCATTTTCTGTCTAAACTTCCTTTAAAGTAATTTTTAATCAGCTGTAAATATCATATCTCCTACTGTTGAAAGTAACTTTAATTTACATTGCACCATATAGCTTGAAAACCAACTTTTGAAATTCTGTACTCCTCCACAAGTGACCTCCGCTAAAATACCCATAGGAAGCTTTACTTTGTGCATGCNTGCTTTGTGTGCCGGTTGCCGTCCTNGGTTGCTTTGGG-3′;5′-TTTTTTTTTTTTTTTTTTAGTGTAAAATACTTAAGCGTTTCCACTA (SEQ ID NO:24)TTGGAAGAAAAGCATATATGGGTATTTTGTAITGTAACTTGTTTAAAAGGACAGTCTTTTTTAATCTTCCCACTTAAATGCTTTTAAAATATGTAATACAATTTGAAGCTTGTTTAAAAATAGAATTAAATGTCTTATATAGTGCTACTGTTTTGGAATTAGAAAGTGATCAAATACAAAACATTTTAAAATTAAGCCCAGAAAACAAAATAGTGTTTAAAGTTAGTTTAGTATAAAAGAAATTTATGAGATTTTTTCTTCAATATAAGATACCTCACTTGAAAATAAAGAAAGCACAGCACATTAAAGTAATTCTCATGAGAACACCCCATTAGAATAATTGCTAAATCTAGGACACCTTTTGAGTTGTGAAGTTTGTGATACATGTAGTCACCATTAGCTTTTCTGCTGGAAGGACTTCCCGTAGTAATTTTAAAGNAGTGTAATAAGTTCAATTANCCCACAAGTTTCTAANCTGGGAAAGNAANTATGGTGAATGGNCCCTTTCTGCAACTACGGGAACACA-3′ ; and 5′-TTTTTTTTTTTTTTTTTTTTGGCATTAA (SEQ ID NO:25)AAGTGTTTATTGGGAATATCATCCAATCTATACAAGTTATATACAAGGCATGAAAATGGCAAACAGCACAAAATACGATTGAGGTATAAGCTAAGAGCACAGTATGTCATGTTTCAATAAATATAATCCAAAATTTGTAAACTAAGTAACCAGATAGATGAGTCATTTTTTTAGTAAAACCATATAAAATATTTATTTCATGTGAGGTAGAGGACAGTTTTGTGTGTCGTGTAATGCAACCAACCACAGCAATTTTAATCATAAAACTATATGCACTGGCAAAATTATCAATCGAGTTATGCTCAATGTAGCTAATGTGTTTCCGTAGTTGCAGAAGGGACCATTCACATACTGCCTTCCCAGGTTAGAAACTGTGGGGTAATTGAACTATTACACTGCCTTAAAATTACTACGGGAAGTCCTTCCAGCAGAAAAGCTAATGGTGACTACATGTATCACAAACTCACAACTCAAAAGGTGTCCTAGATTTAGCAATTATTCTAATGGGGTGTTCTCATGAGAATTACTTTAATGTGCTGTGCTTTCTTTATTTCAAGTGAGGTATCTTATATTGAAGAAAAAATCCATAA-3′

[0155] This clone is similar to sno I.

[0156] Another IEG nucleic acid clone was designated L100. The L100clone is 2924 bp in length and has a nucleic acid sequence as follows:5′-TGCGGCCGCCGGGGCCGGG (SEQ ID NO:26)GCTGAGCCAGTCTCTCCCGCCGCCGCCGGACGCGCAGACCTGGGCAGGCTGCACCGACGGCCGCCTGGCCGAGCGCACTGCAGGTCGCTGCGCGCGCTGCGACCCCGGGGCCCGGACGCGAGTGGCTGCGGTGTCCTGGGCGAGCACTGCTAGTTTAGGCCGTCTGTCCCAGCTGCTTTGGAACCCCTACATCCCACCATGGCTGGGATACAGAAGAGGAAGTTTGACCAGCTGGAAGAGGACGACTGCAGCTCCTCCTCCTTGTCCTCTGGCGATCTCTCTCCCTCTCCTCCCAGCTCTTCTGCCTCCCCTGCCTGGACCTCTGAGGAGGAGGGACTGGGTGATCAGCCACCCCAGCCTGATCAGGACTCCAGTGGCATCCAGAGTTTAACGCCCCCATCCATCCTGAAGCGGGCTCCTCGGGAGCGTCCGGGTCACGTGGCCTTCGATGGCATCACTGTCTACTATTTCCCGCGGTGCCAGGGATTCACCAGTGTGCCCAGCCATGGTGGCTGTACCCTGGGCATGGCTTCTCGTCATAGCACCTGCCGCCTCTTCTCCTTAGCCGAGTTTAAACAGGAGCAGTTCCGGGCTCGGCGTGAGAAGCTCCGTCGGCGTTTAAAGGAGGAGAAGCTAGAGATGCTGAAATGGAAGCTTTCAGTGTCCGGAGTTCCGGAGGCAGGGGCAGACGTGCCGCTCACAGTGGACGCCATCGATGACGCTTCTGTAGAGGAGGACTTGGCAGTGGCCGTGGCAGGTGGCCGCCTGGAGGAAGCGAATTTCCTACAGCCCTATCCACCTCGGCAGCGACGGGCCCTACTTCGCGCTTCCGGTGTTCGAAGGATTGACCGAGAGGAGAAGCACGAGCTGCAGGCGCTACGCCAATCCCGGGAGGATTGTGGTTTGTCACTGTGATGGCGTCTGTGACCCTGAGACCTGCAGTTGCATCCTGGCGGGCATTAAATGCCAGATGGATCACACGTCCTTCCCCTGTGGCTGCTGCAGCGAGGGCTGTGAGAACCCCCATGGTCGAGTGGAATTCAATCAGGCGAGAGTTCAGACACACTTCATCCACACGCTCACCCGCCTGCAGATGGAGCAGGGTGCGGAGAGTTTGGGGGACCCGGAGTCCCCCATGGAGGACGTTCCTGTCGAACAAACCGTGGTTTCCCCCTTTCCTCCTTCCAAACCCACTATGAGCAATGACCTGGGGGACAGCAGCTGTGGCAGCGACATGACAGACTCTTTCCACGACCTACTCCTCTGGCGGCAGTGGCAGCCGCAGCGAGGCTCCGAACCATCTTGCCCACCCCAGCCTGCCAGGTTCCAGCTfCCGGTCTGGCATAGATGAAGACAGCCTGGAACAGATCCTGAATTTCAGTGACTCTGACCTCGGTATTGAGGAAGAAGAGGAGGAGGGAGGGAGTGTGGGCAACTTGGATAACCTCAGCTGTTTTCATTTGGCTGACATCTTTGGTACCGGTGACCCCGGCAGCCTGGCTAGCTGGACACACAGCCAGTTTGGCTCTAGCCTTCCATCGGGCATCCTAGATGAGAATGCCAACCTGGACGCCAGCTGCTTCCTAAGCAGCGGACTCGAAGGGTTGAGAGAAGGTAGCCTCCCCAGCAGTTCTGGGTCCCCTGAGGGGGAAGCCGCCCAGAGCAGCTCCTTGGACCTCAGTTTATCCTCCTGTGACTCCTTTGAGCTTCTCCAATCTCTGCCAGATTATAGTCTGGGGCCTCACTATACTTCCCGAAGGGTATCTGGCAGCCTGGACAGCCTTGAGACCTTCCACCCTTCGCCCAGCTTCTCTCCACCGAGGGATGCCAGCTTCCTGGATTCTCTCATAGGCCTGTCTGAGCCGGTTACAGATGTCCTGGCGCCCCTTCTGGAGAGCCAGTTTGAGGACACTGCTGTGGTGCCTTTGGACCCTGTGCCTGTGTAAGGATTGAGATGACTTTTTCCTGCCCTGAGACCCTGTTGCTGCTTTTTATGTGATCTTGGTGTCCCCCAAGGTCTGTGTATGTAACGGTCTCCCGTGGGCTGGTTCTGCCCCCGTGCCATGTGGGCAATCCTCTATTTTTACAGTAACACTCTAGATTTATTTATTTTTTTATGTTTTTCTGTACTGAAGGGAGGGTGGGAAGGGTATCCCTCTTTCAATGCCTGGCCTCTATGTCCAAACAGAGGTCTCCCACCTCCTACTGTATGCCTGGAGGAGGAAGGGGCGGGGTTCACATCCCCTCTTTCTGTACTGTAAAATGCTCCTTGGTCCAAAGACAGCTGAAAAGCAGGCCTTAGGGTTTCCTGTGGACCGTGGGAGCTAGGTCTTCTGGACTCTGAAGATGTAATTTATTTCTGTAATTTATTTGGGGACTGAGACAGCAGTGGTTGGGCCTCTCTGGCAGGTGGGCGGTGTTGAGGCAAAGTCTTCGGTGTCCCCCGCCGGTCTGGGCTTCGGTGTGGCGTGTAGGTTCGAGCTGAGCAGACGGAGGCTGTGCTTGACCATCGGTGATCAAAACTCCCTCTGCCCCCTGCCCAGACGCTCTAACATGCCCTCTGTCCATTTCCCTCTCCCCAAGGCCATGGGTTATAAAGGCCCTATGTAGGATGGGGAGCCAGAGGCCCTAAGACATGAAGCACACCCCAGATCACTGTCTCTAGCCITTCTGGGCACTGAATCCATCCTGACCCACCACACACCCCCCGGCCAGTTGGCAAGAAAGAGGTGGCTCTTGGGGGCTTTTATGCCCTTCATTAGCTGATGTTGGATTTTATATGCATTTTTATATTGTCTCTAAGTGTCAGAACTATAATTTATTCATTTCTCTGTGTGTGTGTGTGCCAAGAAACGCAGGCTCTGGGCCTGCCTCCTTGCCCAGGAGGCCTTGCCAGCCTGTGTGCTTGTGAGAACACATTGTACCTGAGCTGACAGGTACCAATAAAGACACTCTATTTTTAAAAAAAAAAAAAAAAAAA-3′

[0157] In addition, the L100 clone contains an ORF from basepair 145through basepair 1890. This ORF encodes a polypeptide of 582 amino acidresidues. The translational start site was assigned to the firstmethionine residue in the ORF. The amino acid sequence of the L100polypeptide is as follows: MAGIQKRKFDQLEEDDC (SEQ ID NO:27)SSSSLSSGDLSPSPPSSSASPAWTSEEEGLGDQPPQPDQDSSGIQSLTPPSILKRAPRERPGHVAFDGITVTTTPRCQGFTSVPSHGGCTLGMASRHSTCRLFSLAEFKQEQFRARREKLRRRLKEEKLEMLKWKLSVSGVPEAGADVPLTVDAIDDASVEEDLAVAVAGGRLEEANFLQPTPPRQRRALLRASGVRRIDREEKHELQALRQSREDCGCHCDGVCDPETCSCILAGIKCQMDHTSFPCGCCSEGCENPHGRVEFNQARVQTHFIHTTTRLQMEQGAESLGDPESPMEDVPVEQTVVSPFPPSKPTMSNDLGDSSCGSDMTDSSTTTSSGGSGSRSEAPNHLAHPSLPGSSFRSGIDEDSLEQILNFSDSDLGIEEEEEEGGSVGNLDNLSCFHLADWGTGDPGSLASWTHSQFGSSLPSGILDENANLDASCFLSSGLEGLREGSLPSSSGSPEGEAAQSSSLDLSLSSCDSFELLQSLPDTSLGPHTTSRRVSGSLDSLETTHPSPSFSPPRDASFLDSLIGLSEPVTDVLAPLLESQFEDTAVVPLDPVPV

[0158] This amino acid sequence was found to contain numerous cysteineresidues, forming a motif that has features of a methalothionein-likemotif. Alignment analysis revealed that the L100 methalothionein-likemotif exhibits higher similarity with the methalothionein motif from C.elegans than with the methalothionein motif from mouse.

[0159] Northern blot and in situ analysis using a sequence from the L100clone revealed that L100 mRNA is weakly expressed in wild-type ratbrain. For in situ hybridization, Dig-labeled cRNA probes were used asdescribed elsewhere (Kuner et al., Science 283:5398 (1999)).Specifically, this weak L100 mRNA expression was observed in thepyramidal cell layers as well as the dentate gyrus of the hippocampus,thalamus, cortex, cerebellar granule cell layers, and several fibertracts including the fimbria hippocampus and the cingulum. In addition,Northern blot analysis revealed that the expression of the L100 mRNA wasstrongly upregulated in response to the multiple MECS treatment.Specifically, L100 mRNA expression was induced 17.2 fold by the multipleMECS treatment as determined from Northern blot data using total RNAfrom rat hippocampus (Table I).

[0160] The mRNA expression pattern of L100 demonstrated a compellingoverlap with neuronal populations known to release Zinc into the synapsevia synaptic vesicles and to take-up Zinc post-synaptically. Briefly,synaptic release and uptake of Zinc may participate in the induction andmaintenance of epileptic seizures and the neuronal cell death followingepileptic seizures and ischemia. The L100 metallothionine-like motifmost likely enables the L100 polypeptide to bind Zinc or other divalentcations in vivo. The expression of L100 mRNA in Zinc-containing neuronalpopulations in the brain indicates that L100 polypeptide may sequesterZinc in brain.

[0161] In addition, when acute seizures were induced by kainatetreatment, the expression of L100 mRNA was strongly upregulated (TablesII and III). Kainate-induced seizures is a model used to study epilepsy.Briefly, 300-350 g male Sprague-Dawley rats were intrapertoneallyinjected with either 10 mg/kg body weight of kainate or PBS. RNA samplesfrom the hippocampus, cortex, and cerebellum were prepared from treatedrats at 1.5, 6, and 24 hours post-injection. This RNA then was used tomeasure mRNA expression by Northern blot and RT-PCR analysis. ControlmRNA measurements included c-fos, GAPDH, NO-38, and ATF-4 for theNorthern blot analysis, and Hsp70, c-jun, Zif268, c-fos, Clathrin, andβ-actin for the RT-PCR analysis. A Phosphoimager FLA2000 (Fuji) was usedto analyze the data, which was expressed as the Integral PSL-backgroundPSL (ID evaluation with Aida version 2.0).

[0162] At six hours following kainate injection, strong upregulation ofthe L100 mRNA was observed, by in situ hybridization, in the dentategyrus and areas CA3 and CA4 of the hippocampus as well as the associatedentorrhinal cortex, the cingulum, and fimbria, which are brain areasknown to be highly excited in and which mediate Kainate-inducedseizures. Moderate upregulation of the L100 mRNA also was found in thethalamic nuclei, temporal, parietal, frontal, medial orbital, andcingulate cortex as well as in the cerebellar granule cells. Thus, thedata presented herein indicates that L100 participates in cellularmechanisms mediating kainate-induced epileptic seizures and theconsequent neurodegeneration. TABLE II mRNA expression normalized toGADPH expression 1.5 hour 1.5 hour 6 hour 6 hour 24 hour 24 hour ClonePBS kainate PBS kainate PBS kainate Hippocampus: L100 4622 85251 784715444 3940 16551 L119 2816 69982 4597 11519 2787 12944 Cortex: L100 — —81 290 86 131 L119 — — 255 1262 538 505

[0163] TABLE III Fold increase in mRNA expression upon kainate treatmentHippocampus Cortex Clone 1.5 hour 6 hour 24 hour 1.5 hour 6 hour 24 hourA013 9.8 — — L094 3.6 — — L100 18.44 1.97 4.20 3.58 1.52 L119 24.85 2.514.64 — R113 2.0 — — R286 — — —

[0164] In addition, when acute seizures were induced bypentylenetetrazole (PTZ) treatment, the expression of L100 mRNA wasstrongly upregulated (Tables IV and V). PTZ-induced seizures is a modelused to study epilepsy and ischemia. Briefly, 300-350 g maleSprague-Dawley rats were intrapertoneally injected with either 50 mg/kgbody weight of PTZ or PBS. Total RNA samples from the hippocampus,cortex, and cerebellum were prepared from treated rats at 20 minutes, 6hours, and 24 hours post-injection. This RNA then was used to measuremRNA expression by Northern blot analysis. Control mRNA measurementsincluded c-fos and GAPDH. A Phosphoimager FLA2000 (Fuji) was used toanalyze the data, which was expressed as the Integral PSL-background PSL(ID evaluation with Aida version 2.0). TABLE IV mRNA expressionnormalized to GADPH expression 20 min 20 min 6 hour 6 hour 24 hour 24hour Clone PBS PTZ PBS PTZ PBS PTZ Hippocampus: L100 534 1637  854 1992 966 1903 L119 342 965 — — — — Cortex: L100 958 2719 1162 3740 1175 1825L119 577 1605 — — — —

[0165] TABLE V Fold increase in mRiNA expression upon PTZ treatmentHippocampus Cortex Clone 20 min 6 hour 24 hour 20 min 6 hour 24 hourL100 3.1 2.33 1.97 2.84 3.22 1.55 L119 2.82 — — 2.78 — — R113 — 2.0 —R286 — 2.6 —

[0166] In another study, the expression pattern of L100 and L119 wasdetermined using two models for ischemia. Briefly, neurons degenerate inbrain and spinal cord after acute insults (e.g., stroke, cardiac arrest,and trauma) and during progressive, adult-onset diseases (e.g.,amyotrophic lateral sclerosis, and Alzheimer's disease). Impaired energymetabolism plays an important role in neuronal cell death after brainischemia, and apoptosis has been implicated in cell death induced bymetabolic impairment. The irreversible inhibitor of succinatedehydrogenase in the mitochondria, 3-nitroproplonic acid (3-NP),inhibits oxidative phosphorylation and causes intracellular hypoxia.Thus, one model used to study ischemia involves intrastriatal injectionsof 3-NP, which is known to produce selective cell death similar to thatobserved in transient ischemia and Huntington's disease (McLaughlin etal., J. Neurochem 70:2406-2415 (1998)). The other model is a globalischemic paradigm that involves a 15 minute insult by complete occlusionof the carotis.

[0167] In the 3-NP study, 220-300 g Wistar rats were intraperitoneallyinjected with 20 mg/kg body weight. Three hours post-injections, thebrain was removed and total RNA prepared. In the global ischemia study,220-300 g Wistar rats were received a 15 minute insult (bilateralocclusion of the Carotis/arterial pressure=35 mm Hg). One hour later,the rats received a reperfusion followed by immediate brain dissectionand total RNA preparation. Untreated rats were used as controls for eachstudy. Ten (10) μg of total rat brain RNA (without cerebellum) wasloaded per lane and blotted. Probes were prepared from the 3′untranslated regions of L100 and L119. The Northern blot data wascollected using a Phosphoimager (FLA2000 Fuji, Tina software) andexpressed as PSL-background.

[0168] L119 mRNA expression was upregulated 6-fold by global ischemiawhile L100 mRNA expression was not inducible by global ischemia (TableVI). This result indicates that only seizure related stimuli alter theexpression level of L100 and that L100 is not a general marker forstress response of the cell like c-fos. TABLE VI mRNA expression after3-NP or global ischemia treatment. Probe Untreated 3-NP Global Ischemiac-fos 18.1 26.64 216.22 GAPDH 487.02 587.51 593.31 L100 30.95 43.8240.15 L119 55.48 41.94 332.73

[0169] Northern blot analysis using multiple tissues from rat revealedthat the expression of L100 and L119 mRNA was not brain specific (TableVII). Briefly, fragments from the 3′ untranslated region of L100 andother IEG clones were labeled with ³²P-dCTP. The denatured probe washybridized with 10 μg total RNA from rat brain, liver, lung, muscle,intestine, eye, heart, testis, and kidney in the Quik Hyb-solution(Stratagene) at 68° C. and washed with 0.1×SSC at 60° C. For L100, afterone day of exposure, signals were detected at the 3 kb position inbrain. In addition, a weaker signal was detected in heart and a faintsignal detected in kidney. A strong signal was detected in testis butthis signal was at a position corresponding to a size smaller than 3 kb.For L119, a strong signal was detected in heart and weaker signal inbrain. In addition, only very faint signals were detected in liver,kidney, and testis. TABLE VII mRNA expression in various rat tissues.Probe Brain Liver Lung Heart Kidney Muscle Intestine Testis Eye A013 (+)(+) (+) (+) L094 + + (+) + (+) + L100 +++ ++ + +++(*) L119 ++ +++ R113(+) (+) (+) (+) (+) (+) (+) R286 +++ (+) +++ (+) + (+) (+) ++

[0170] Another IEG nucleic acid clone was designated L111. The firstround of screening produced a clone (designated L111-5) that contained a3.0 kb fragment of L111. A second round of screening using the codingregion of L111-5 as a probe produced several additional clones. Thefollowing nucleic acid sequence is within the L111 clone:5′-ATTCGGCACGAGCCAGAG (SEQ ID NO:28)TGAAGGGGCATGGAGAAGTGGACGGCCTGGGAGCCGCAGGGCGCCGATGCGCTGCGGCGCTTTCAAGGGTTGCTGCTGGACCGCCGCGGCCGGCTGCACTGCCAAGTGTTGCGCCTGCGCGAAGTGGCCCGGAGGCTCGAGCGTCTACGGAGGCGCTCCGGCAGCCAACGTAGCTGGCAGCTCTCTGAGCGCTGCTGGCGCCCTAGCAGCCATCGTGGGGTTATCACTCAGCCCGGTCACCCTGGGAGCCTCGCTCGTGGCGTCCGCCGTGGGCTTAGGGGTGGCCACCGCCGGAGGGGCAGTCACCATCACGTCCGACCTCTCTCTGATCTTCTGCAATTCCCGGGAGGTACGGAGGGTGCAAGAGATCGCCGCCACCTGCCAGGACCAGATGCGCGAACTCCTGAGCTGCCTTGAGTTCTTCTGTCAGTGGCAGGGGCGCGGGGACCGCCAGCTGCTGCAGAGCGGGAGGGACGCCTCCATGGCTCTTTACAACTCTGTCTACTTCATCGTCTTCTTCGGCTCGCGTGGCTTCCTCATCCCCAGGCGTGCGGAGGGGGCCACCAAAGTCAGCCAGGCCGTGCTGAAGGCCAAGATTCAGAAACTGTCTGAGAGCCTGGAGTCCTGCACTGGTGCCCTGGATGAACTTTAGTGAGCAGCTGGAATCCCGGGTCCAGCTCTGTACCAAGGCCGGCCGTGGTCACAACCTCAGGAACTCCCCTGATCTGGATGCAGCGTTGTTTTTCTAAGAGCATCCTCTAGCTGTGTGGAATGTTCTAGATTCGCAGCATCCACAAGGAAGTGCTACATGGGCGGAGTGCAAAGGATTICAGAAGCTCTTCTTGCAGGGCATCAGTCCGTAGCTCCTTGTGTGTGCGAAAGACTTTTCACTTGTGTAATCCCAACTGAGTATGTGACCCTAAACAGTCACTTTGGGGACTCCCCAAATCCTTTTTAGCTGCACACAGCTTGTCAGACTGTCCTICAATTAGAGTTATTGGGGTGGGGGGGCTTGATGGCTTGAGTAATAGAGGTCTGGCGAGGTGTCTCCCTCTTGGACCTCTTATGTGTTGTTACTAGAATCCTGAGATTCTCAAATGTTGGTGAGAGGAGACTTTTACTTTTCAACTTTGCTTCGGCAGTTTCCGATACACAGGACTCCAGAATCCAGAACAAGAAAGAAGAACCTTGTGTTTGTAGGGTGTGCAGACCCAGACGGGGCCGAGGAGCTGACTTGCTCAGCTCTCACACGCAGCCAGTTTATCCACTCACAGACCAAACCTGGCTACTGCATAGACTGTTCCAGTGTGGCTTCAAATCCACACCTCTAGGTACCCTGAGAAGGAAAGCCACCTGAAGAGTCACTCTAATCCCAACACGCTCACCCCCTTCACGTCCATAAAGGAGCTGGGCAAGGGGTGAGATGAAGACCCTGACAATTTTAAATGACTGTAGCATAGAGAGCCATGGCCTTTGAGTTTAAGAGTCTTGATCCCAGGTTCTGTCCCCCACTGTCCTGTGACTTAGCCACCTTGTCTTGCTACAGATGGTGGTAGGAGGCCACCCTGTTGCGAAGTCCTGAGATAATGACAAACACAGAGGCTAGCTCACAAAAATGTACTTCCTGGCCTGGCTTCTGAAGGGTTAACTGTTGGGCTCCATCCCAGATTTCTGAGATCAGGAACTCCAAATATGAGGCCCGCCTCTGGCTGATTCTGATGCCCCATAAATGTTTGAAAATGACACAGCAAAGGTTCATCTCCAGCCAGGTGTGGTGGGACACACCTGTAAGGCCAGCGCTTGGAGATGGAGACAGGGGGACCAGTAGTTCAGGGTCATTCTTGGCTACATAGCAAACTCAAGGCCACCCTGGTCTCAAAAACCAAAACAAAAAGCCATCTTCTGACTCCCTTCAATTGTTCAAAGCCTTTCCAGGGCCTTCAGAATCACGCTCAGAGTGTTCTGGGAAGATTAGCCCAGAAGCCAGAGAAAGAGTACGCTGTGTGCTTGTAAAGCCAGTTACTCTGTCCCCTGTGAACTAGGAGACAGAGCACTTCCGACCCTATAGAGGGCAGTAGTGGCCATTCCTTGTAGGGGACTGGTATAGAAGTAATGTGAACTATTTAAAAATAGTTATTTAATTGCTGCCTTCACATTTGATTTTATTTAACCTTTTCACATTATTTAGAAAATAATAAGAGTAGTAAGTGTCTGAATAGGAAGGGAGTCTCTTAAGGCTCTTTCCAAGAGCTCAGGTTTGGATTTCTAGAGTCCCCCCGACCCCAGAGAGGACTCTTTAGTGTTTGACACGGTCTTTGTAAGTAAGATGGGGAGTCCTGGAGAGAGAGACCAAGCTGATTTTTAAACTAGGAAATGGAGTCTTGAACTGTGGAAGATTTGAAAAGTTAAGCCTATGTGTCTTGAAGGTACTTGGCCAGAAAAGCACTTGGCTTGAAAAAGAAAACCTGTTTTAATTCAGGGGTGGAGGAATAGAGACAGATGAAGAAAGCATTTAGACCTCGGAAACCTGATGTCCTATGAAATTCTGTTTTTATAAAATTGTGTTATGGTGGAGATCTGTTGCATTTCGACTTTGTGGCTGTAAGAAACCTGTTATCTATGTTTAAGAAAGTACTICTAATTTATTCAATGTCTTCCTAAATTATCCTTTAAAAAAAAAAGTTGGAAAGTCTATGAGACCGTACCTAAGAAACCTTGACTGTGTATTTTAAGTTATTTAATGCCATGCATTTGTGAAGCCCCTTCCCAGTGATGGCTGTGGTGTGTCTGAGGAAATGTAAGTTTGGCATGAGGGGGAGGGGCTGCTGTTTCTATATTTGTTTTTGTTTTCTATAAACAGTAATCAGGATGTATCCTGGTTTCATTTGACATTGAAAAAAAAAAAAAAACTCGTGCCGAATTC-3′.

[0171] The L111-5 clone contained 0.5 kb of the 3′-end of an ORF.

[0172] Northern blot analysis using a sequence from L111 revealed thepresence of a 4.0 kb mRNA transcript. This analysis also revealed thatthe expression of L111 mRNA was marginally upregulated in response tothe multiple MECS treatment.

[0173] Another nucleic acid clone was designated L117. The L117 clone is2460 bp in length and has a nucleic acid sequence as follows:5′-TACGGCTGCGAGAAGACGACAG (SEQ ID NO:29)AAGGGGAGCGGAGCCAAGATGGCGGCGGAGCTGGAATACGAGTCTGTGCTGTGTGTGAAGCCCGACGTCAGCGTCTACCGGATTCCGCCGCGGGCCTCCAACCGCGGTTACAGGGCATCTGACTGGAAGCTAGACCAGCCTGATTGGACTGGTCGCCTCCGAATCACTTCAAAAGGGAAGATTGCCTACATCAAACTGGAAGATAAAGTTTCAGGGGAGCTCTTTCGCTCAGGCGCCAGTAGAGCAGTACCCTGGGATTGCTGTGGAGACTGTGGCCGACTCCAGCCGCTACTTTGTGATCAGGATCCAGGATGGCACCGGGCGCAGTGCGTTTATTGGCATCGGCTTCACGGACCGGGGAGATGCCTTCGACTTTAATGTCTCCCTGCAAGATCACTTCAAGTGGGTAAAGCAGGAAACCGAGATCTCCAAAGAATCGCAGGAAATGGATAGTCGTCCCAAGTTGGATTTAGGCTTCAAGGAAGGGCAAACCATCAAGCTGAGTATTGGGAACATTACAGCCAAGAAAGGGGGTACTTCTAAGCCCCGGGCCTCAGGAACGGGGGGCCTGAGCTTACTCCCACCTCCTCCTGGAGGCAAAGTCACTATCCCCCCACCGTCCTCCTCCGTTGCCATCAGCAACCACGTCACCCCACCACCCATTCCAAAATCTAACCATGGAAGTAATGATTCAGATATCCTGTTAGATTTGGATTCTCCAGCTCCTGTCCCGACTTCAGCACCAGCTCCAGCTCCAGCTTCTACAAGCAATGACTTGTGGGGAGACTTTAGCACTGCATCCAGCTCTGTTCCAAACCAGGCACCACAGCCATCTAACTGGGTCCAGTTTTGAGTCGCATTGGCAAGAAGTTGAGGACACTTGAAGAATAAAAATGACCTCAAGGGCACCATTCTATGAGGGAGTTGAGGGACGGCTTAATTTCCCAGGACCCAAATCAGTGGTCAGTCTTTCCTGTAGCTTCTCTGTGCATTCAGGCTGGATTTTTTTTTTTTTTTTTTTTGGTTACCTCTGTGTTACTTGCTGTATATCCAGGAGACAATCTGCTGTTTCCTGCTCAGAACCAAGCAAGGGAGTAGTGGGTATTATCACACTGACTGACTTTGCAGAGTTCAGAAGGCCAACTTGATGAGTGGGAGTGACCTCGAACGTATGTAAATCCTTGAACTTATTTCAGAATCATCTCATGATTCCCTAGTTAGCAATTTCAGGAGAGACAAATGCCTTGAAACTGTCTTCTCCACTAATCCGAGACTAAATATGGTCAGGCTGGCCCCAGGACTCATGAAGTTAGGGTTTTCATGGGGGTAGATTTGGAGAAAGCTGTGTCCCGGCTCTCTTCTGTAAGGCCTCCTTCAGGCTTACCCCATGCAGTGAACTTCCCGTGCTGGGTGGAGCCCCATCACCTTCTTTGTGTGTTTACATGTTGTTTCCTTTGACAAGAGGGTTATGTTGGTGGCACCTCACTGTTTTCTTGTTGAATAGTGCAGCATCTTTGACCAGTGAATATTTCTGAGATGAAGGGGTCAAGGGGCTGTGCTTTCCATGGTGTAGTCTACAGAAGTGTTTAATTTCTTGCGGCCCCACGGGATTGCTGCACTGACGCATAGAATTGATCTATACTCACCCTGTGTTTGACCTGAAGAGTTTTAACTTGATGTGTAGAGCAGAGAGCTGGAAGCACTAAGTTCCCATTCAGTACCCACAATGCCTTGCTGCCTGGTTTGACTCCTTTTCATAAACATTTCATTTCAGTCCATCTAGCACTTCTGTGGAAAGCTGCTGTTGATTGTGTCAGTGTGAAGGAGGTGAAGTCACAGCTTTCTTTACCTATGACAGTTAGGCTTTGCACTAGACGTTTGATACCAGCTAGGATATCTTAAAGGAAGTTACCGCCCCATCACTCTCCAGTCTCTGGCCGCCATTCCTTTTACAGTGCTGTGAAGAGCGTCCTCTGAGGTCGGTGGGTACTGTCTCCTGTTGGTCGGGCAGTTTGAGGGAGGAGTGGGAGGACTCACACTCCTGCAGGTACCTGTTTGGGTAGCACACTGGCTGCAGAGAGTCCTTTCAGATATATTGTTTCTCAATGTTCTTCGTAGCTTTTTCTAACTTCGGGTCCATTTTTCCCATCGCCTCTTCCCATTCCCAGGCAGCTCTCTTGTTGCAGAGCCATGGCAGGACGTTTAAGTTTCCAATAAAAACACTAAGAAGAAAGTATAGAATCACTAGTGACTGTTGGGAAACCTATTTTCTCAATCTTCCTCCATTTTGTGTTCTTTGTATTCTTAAGATGATAATATATTATGTATTTGAATTGCTGAAAATTGAAAATGAAGTTGAAGATATATGTATATAAGCGTATGCTGTAT1GGTGCAATAATGGTAATTAAAGATATTAAAAAAGAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAA-3′

[0174] In addition, the L117 clone contains an open reading frame (ORF)from basepair 42 through basepair 875. This ORF encodes a polypeptide of278 amino acid residues. The amino acid sequence of the L117 polypeptideis as follows: MAAELEYESVLCVKPDV (SEQ ID NO:30)SVYRIPPRASNRGYRASDWKLDQPDWTGRLRITSKGKIAYIKLEDKVSGELFAQAPVEQYPGIAVETVADSSRYFVIRIQDGTGRSAFIGIGFTDRGDAFDFNVSLQDHFKWVKQETEISKESQEMDSRPKLDLGFKEGQTIKLSIGNITAKKGGTSKPRASGTGGLSLLPPPPGGKVTIPPPSSSVAISNHVTPPPIPKSNHGSNDSDILLDLDSPAPVPTSAPAPAPASTSNDLWGDFSTASSSVPNQAPQPSNWVQF.

[0175] Using tblast2× algorithms, the L117 polypeptide was found to havehomology with expressed sequence tags (ESTs) from mouse, mouse embryo,human hNT neurons, human tumors, drosophilia, drosophilia embryo, C.elegans, and Arabidopsis thaliana, a plant organism. Although thesequence of ESTs can be questionable, the identified ESTs were alignedfor comparison. The comparison of consensus sequences from each speciesprovided evidence that the L117 clone or a L117 motif has a very strongpressure for conservation during evolution since it is conserved in avariety of very distant species. In addition, this alignment indicatedthat the first methionine residue in the ORF of the L117 clone is thetrue initiation site for translation since most of the homology betweenthe ESTs begins around this position, and the C. elegans, drosophilia,and human hNT ESTs each contain a methionine residue that is in a veryclose proximity to that of the L117 clone. Further, the relation betweenthese ESTs and the L117 clone was supported by an exactly matching stopcodon in the human EST, mouse EST, and L117.

[0176] Northern blot analysis revealed that the expression of the L117mRNA was not upregulated in response to the multiple MECS treatment ineither the hippocampus or cortex. Analysis using a total RNA extract,however, revealed a small upregulation upon MECS stimulus.

[0177] Another IEG nucleic acid clone was designated L119. The L119clone is 2900 bp in length and has a nucleic acid sequence as follows:5′-ATTCGGCACGAGCCAGAG (SEQ ID NO:31)TGAAGGGGCATGGAGAAGTGGACGGCCTGGGAGCCGCAGGGCGCCGATGCGCTGCGGCGCTTTCAAGGGTTGCTGCTGGACCGCCGCGGCCGGCTGCACTGCCAAGTGTTGCGCCTGCGCGAAGTGGCCCGGAGGCTCGAGCGTCTACGGAGGCGCTCCTTGGCAGCCAACGTAGCTGGCAGCTCTCTGAGCGCTGCTGGCGCCCTAGCAGCCATCGTGGGGTTATCACTCAGCCCGGTCACCCTGGGAGCCTCGCTCGTGGCGTCCGCCGTGGGCTTAGGGGTGGCCACCGCCGGAGGGGCAGTCACCATCACGTCCGACCTCTCTCTGATCTTCTGCAATTCCCGGGAGGTACGGAGGGTGCAAGAGATCGCCGCCACCTGCCAGGACCAGATGCGCGAACTCCTGAGCTGCCTTGAGTTCTTCTGTCAGTGGCAGGGGCGCGGGGACCGCCAGCTGCTGCAGAGCGGGAGGGACGCCTCCATGGCTCTTTACAACTCTGTCTACTTCATCGTCTTCTTCGGCTCGCGTGGCTTCCTCATCCCCAGGCGTGCGGAGGGGGCCACCAAAGTCAGCCAGGCCGTGCTGAAGGCCAAGATTCAGAAACTGTCTGAGAGCCTGGAGTCCTGCACTGGTGCCCTGGATGAACTTAGTGAGCAGCTGGAATCCCGGGTCCAGCTCTGTACCAAGGCCGGCCGTGGTCACAACCTCAGGAACTCCCCTGATCTGGATGCAGCGTTGTTTTTCTAAGAGCATCCTCTAGCTGTGTGGAATGTTCTAGATTCGCAGCATCCACAAGGAAGTGCTACATGGGCGGAGTGCAAAGGATTTCAGAAGCTCTTCTTGCAGGGCATCAGTCCGTAGCTCCTTGTGTGTGCGAAAGACTTTTCACTTGTGTAATCCCAACTGAGTATGTGACCCTAAACAGTCACTTTGGGGACTCCCCAAATCCTTTTTAGCTGCACACAGCTTGTCAGACTGTCCTTCAATTAGAGTTATTGGGGTGGGGGGGCTTGATGGCTTGAGTAATAGAGGTCTGGCGAGGTGTCTCCCTCTTGGACCTCTTATGTGTTGTTACTAGAATCCTGAGATTCTCAAATGTTGGTGAGAGGAGACTTTTACTTTTCAACTTTGCTTCGGCAGTTTCCGATACACAGGACTCCAGAATCCAGAACAAGAAAGAAGAACCTTGTGTTTGTAGGGTGTGCAGACCCAGACGGGGCCGAGGAGCTGACTTGCTCAGCTCTCACACGCAGCCAGTTTATCCACTCACAGACCAAACCTGGCTACTGCATAGACTGTTCCAGTGTGGCTTCAAATCCACACCTCTAGGTACCCTGAGAAGGAAAGCCACCTGAAGAGTCACTCTAATCCCAACACGCTCACCCCCTTCACGTCCATAAAGGAGCTGGGCAAGGGGTGAGATGAAGACCCTGACAATTTTAAATGACTGTAGCATAGAGAGCCATGGCCTTTGAGTTTAAGAGTCTTGATCCCAGGTTCTGTCCCCCACTGTCCTGTGACTTAGCCACCTTGTCTTGCTACAGATGGTGGTAGGAGGCCACCCTGTTGCGAAGTCCTGAGATAATGACAAACACAGAGGCTAGCTCACAAAAATGTACTTCCTGGCCTGGCTTCTGAAGGGTTAACTGTTGGGCTCCATCCCAGATTTCTGAGATCAGGAACTCCAAATATGAGGCCCGCCTCTGGCTGATTCTGATGCCCCATAAATGTTTGAAAATGACACAGCAAAGGTTCATCTCCAGCCAGGTGTGGTGGGACACACCTGTAAGGCCAGCGCTTGGAGATGGAGACAGGGGGACCAGTAGTTCAGGGTCATTCTTGGCTACATAGCAAACTCAAGGCCACCCTGGTCTCAAAAACCAAAACAAAAAGCCATCTTCTGACTCCCTTCAATTGTTCAAAGCCTTTCCAGGGCCTTCAGAATCACGCTCAGAGTGTTCTGGGAAGATTAGCCCAGAAGCCAGAGAAAGAGTACGCTGTGTGCTTGTAAAGCCAGTTACTCTGTCCCCTGTGAACTAGGAGACAGAGCACTTCCGACCCTATAGAGGGCAGTAGTGGCCATTCCTTGTAGGGGACTGGTATAGAAGTAATGTGAACTATTTAAAAATAGTTATTTAATTGCTGCCTTCACATTTGATTTTATTTAACCTTCACATTATTTAGAAAATAATAAGAGTAGTAAGTGTCTGAATAGGAAGGGAGTCTCTTAAGGCTCTTTCCAAGAGCTCAGGTTTGGATTTCTAGAGTCCCCCCGACCCCAGAGAGGACTCTTTAGTGTTTGACACGGTCTTTGTAAGTAAGATGGGGAGTCCTGGAGAGAGAGACCAAGCTGATTTTTAAACTAGGAAATGGAGTCTTGAACTGTGGAAGATTTGAAAAGTTAAGCCTATGTGTCTTGAAGGTACTTGGCCAGAAAAGCACTTGGCTTGAAAAAGAAAACCTGTTTAATTCAGGGGTGGAGGAATAGAGACAGATGAAGAAAGCATTTAGACCTCGGAAACCTGATGTCCTATGAAATTCTGTTTTTATAAAATTGTGTTATGGTGGAGATCTGTTGCATTTCGACTTTGTGGCTGTAAGAAACCTGTTATCTATGTTTAAGAAAGTACTTCTAATTTATTCAATGTCTTCCTAAATTATCCTTTAAAAAAAAAAGTTGGAAAGTCTATGAGACCGTACCTAAGAAACCTTGACTGTGTATTTAAGTTATTTAATGCCATGCATTTGTGAAGCCCCTTCCCAGTGATGGCTGTGGTGTGTCTGAGGAAATGTAAGTTTGGCATGAGGGGGAGGGGCTGCTGTTTCTATATTTGTTTTTGTTTTCTATAAACAGTAATCAGGATGTATCCTGGTTTCATTTGACATTGAAAAAAAAAAAAAA A-3′.

[0178] In addition, the L119 clone contains an ORF from basepair 28through basepair 768. This ORF encodes a polypeptide of 247 amino acidresidues. The translational start site was assigned to the firstmethionine residue in the ORF. The amino acid sequence of the L119polypeptide is as follows: MEKWTAWEPQGADALRRFQGLLLDRRGRLH (SEQ ID NO:32)CQVLRLREVARRLERLRRRSLAANVAGSSLSAAGALAAIVGLSLSPVTLGASLVASAVGLGVATAGGAVTITSDLSLIFCNSREVRRVQEIAATCQDQMRELLSCLEFFCQWQGRGDRQLLQSGRDASMALYNSVYFIVFFGSRGFLIPRRAEGATKVSQAVLKAKIQKLSESLESCTGALDELSEQLESRVQLCTKAGRGHNLRNSPDL DAALFF.

[0179] Hydropathy plot analysis revealed a stretch of about 50hydrophobic amino acid residues, possibly indicating that the L119polypeptide is a type II transmembrane protein.

[0180] Northern blot analysis using a sequence from the L119 clonerevealed that the expression of the L119 mRNA was strongly upregulatedin response to the multiple MECS treatment. Specifically, L119 mRNAexpression was induced 17.8 fold by the multiple MECS treatment asdetermined from Northern blot data using total RNA from rat hippocampus(Table I).

[0181] Another IEG nucleic acid clone was designated R010. The R010clone is 1280 bp in length and has the following nucleic acid sequence:5′-GCTTTGGAAACCGGACTGCAGGCT (SEQ ID NO:33)AAACTGGCTTCTTTTGAATCCTTGGAAGCATAAAGGACAAGTAGCAGGGCTCGCAGTCTTCCATTTGTCACTGGAGAAGAACTTATAATTCAGAAGATCTGGGTCTGGACCCAGGCTGACCACTTTGGAGCTTTGAGACTCTGGGATTGTGATCCAGTTCTGAGCTGGTGATAAACACTCCTTGTGACTTTTGGTCAATTCAGCTACCAGATTCCAGCCAACATGACCCTCGCAGCCTATAAGGAGAAGATGAAGGAACTCCCACTAGTGTCTCTGTTCTGCTCCTGTTTTCTGTCTGATCCCCTGAATAAATCATCCTACAAATATGAAGGCTGGTGTGGGAGACAGTGTAGGAGGAAAGGTCAAAGCCAGCGGAAAGGCAGTGCTGACTGGAGAGAAAGAAGAGAACAGGCAGATACGGTAGACCTGAACTGGTGTGTCATCTCTGATATGGAAGTCATCGAGCTGAATAAGTGTACCTCGGGCCAGTCCTTTGAAGTCATCCTGAAGCACCTTCCTTTGACGGGGTGCCTGAGTTTAATGCCTCCCTCCCAAGACGTCGAGACCCATCGCTAGAAGAGATACAGAAGAAGCTAGAAGCAGCAGAGGAGCGAAGGAAGTACCAGGAAGCTGAGCTCCTAAAACACCTTGCAGAGAAACGAGAGCATGAGCGTGAGGTAATCCAGAAAGCTATCGAGGAAAACAACAACTTCATCAAGATGGCGAAAGAGAAGCTGGCCCAGAAGATGGAGTCCAATAAGGAAAACCGGGAGGCCCATCTGGCTGCCATGTTGGAGCGGCTGCAAGAGAAGGACAAGCACGCAGAGGAGGTGCGGAAAAACAAGGAGCTGAAGGAAGAGGCCTCCAGGTAAAGCCCANAGGCCAAGGAAGTTTCCAGGACAGCCGGACAGCTCCCGCAGCAACCTGGTTCCAGCAGCATCGGCCGCTGGCTGCTCTCCCAGCACTGGGGTTCGGGGGGAGGGGGGTGGCCAAAGGGGCGTFTCCTCTGCTTTTGGTGTTTGTACATGTAAAAGATTGACCAGTGAAGCCATCCTATTTGTTTCTGGGGAACAATGATGGGGTGGGAGAGGGGACAGAGAGTGTTTGGAAAAGGAGGTGAAGATGAGCCCGAGGACTTTGTGACACTGTCCACTGACTGCAGACTTGGGCCAAGGCCCCCGCTTTTCACGGCTCTGCCTGGACATTCGGCCTCCAGGTTCCTAGTGGAGAGAAGATGTGACAGAAGTTCAGAGTGAAGGGCCGAGTCCTGGTGGGGTGGTGTGCAGGGCCAGCAGGACGAGCCCGTCTGGATGGAGTGAAACCTACCCTGAGCGGGTGGGATAAGGTCTGTGTGCGTCTGTTCATTGTCATCTTTTGATCATCATGACCAACGAAACATTTAAAAAAAAAAAAAAAAAAAAAA-3′.

[0182] Two genomic R010 clones were also obtained. The nucleic acidsequence for these genomic R010 clones is as follows: 5′-GATAA (SEQ IDNO:35) ACACTCCTTGTGACTTTTGGTCAATTCAGCTACCAGATTCCAGCCAACATGACCCTCGCAGGTAGGTACATGCACCAGTCAGTGATGAACACCATAACACAAGCCATTTTTCTATCTCTGTGTGTGTCCATGTGTATTAAGGTGCATCCGTGTGTGTGATACACACGTAGGTGCATGGCATGCATGTGTGTGCAAATGCATATACAAGTCCAAGGACAGGGGTTGGGGATTTAGCTCANTGGTAGAGCACTTGCCTANGAAGCGCAAGGCCCTGGGTTCGGTCCCCAGCTCCGAAAAAAAGAACCAAAAAAAAAAAAAAAAAAAAAAAAAAAATTTCCANGGACAACCCCAAATTTCCTTTCNCNAAANCCANCCANCTTCCATTNAAAAAAAANGGGTCNCNCNTGGGTfAAACCATTTNNAAANGGCNAACCTNACNGGCCAKTGAKTGCCAGGAATCTTCTTATYCCTGCCCWACCTCCAATGTCTTTCACATGTGAATGCTGAGGGTCAGAACTTGTGCTTACAAGGCAGACATTTTGCCAGCTCTCCGGCCATCTTTCTCTATGTATGTACACTCACAGATGCACAGGAAGAGAGGGTAGAGAAGCCAAGAGGCAAAGTCATTTCTGGGTGGTGGGTGGGATCACAGCTGAATTCTTCTTCCTCATTTGCTCTGTGTGTATTATTTAATTTTAAAATAATACCTTTATAATAGTATCGAAACTATGCTTTCAAGTTTGTAAGAGAAAGTGATCACTGGGCTGTGTAGTGAGGGGGTCTTTATATTATGCATATAACATGGTGCAATGGGAAGGACTGGCAGAGGCCTCCATGATGACCTATGACTTCTAGGGAGACTCAGTCGTGTCAAGGGTACATTCCTACTCTGCAGACAGCTTCTCCCTGGTTTGATTCCTGTGCTGGGAAGATTTGAGGAGTCTTCCAGCCTGACCTCTTCTACAGTGGGCCTGGACTTTAAGGAGAGTAGCAAGGAAGTCTTTTTATTAATCTCTTACCCTTTAGGCAGCAGTGTCAAGTACTTTTAGCAGAATTAAATATAGATTTCCTACAAACTACAAACTTCAAAGCCCTGGTTTATCCTTGGGTGGGAGTAGGAGATGGAGGGCCAGGGTCAGGGCACTGCACTTGGGATCTTTACTTGAGGGTACTCAACGCTTGGTAGTAACAAAAAGTGGGGTGAGTGACAATGTTAATTTTCAACTGGGAGGTAGCCCAGGCTTGGGTACTTTGGAGCCAGAAAGCCTGGGCTGACTCACAGAAGTGGTGCTCTCTCTYGYAGCCTATAAGGAGWWGATGAAGGAACTCCCACTAGTGTCTCTGTTCTGCTCCTGTTTTCTGTCTGATCCCCYGRATAAATCATCCTACAAATATGAAGGTGAGTAGGGGCTAGGCTGGGATAGAAAAGGGTGGAGGCTTCTGTGTCCTGTGTTTGTSGGTGCCCCACATTGACTCCTATCTTGTAAAACTGTCCTGGTCGCAGTGTGTCTTATTTCCCAGAGGCTGAGGAGTCTGAGCCCAGGGGGATGTAGCCTGGGTGCCAAGCAGCCTCCAGGGATCTGGATTGGGCCCTCCTGGAGCACTTGCTCCTAGAGTCCCTTTTRCACATTCCTTGACACCACAGAGGACACCAGGATAAGCCAGACACAAGTTTTGAGATTCCATTCATGGAGGCCCAGAACAGAAAAAGAAAACTTAGTGTGTTCACCAGGGCTTCTAGGGACAGGTAGAGATGCTCCTAGACAGGTCCAGGGTGGGAATAGCACTTCTAACCTGGATGGTGACAGTTCGAGCCCCTAGACCCTATCAGAGAGTACTGGATTGTCATGCTGTCAGGAGGAGTGGTCAGGGGACAGATAGGTCATCTCTTCATTTCTGTTTGCCAGGAAGGGATGGGTTTGGTCTGTCAATAAGAGAGATGGGTGTTTGGATGACCTGAGTCTGTTTTTTCCATTTAGGCTGGTGTGGGAGACAGTGTAGGAGGAAAGGTCAAAGCCAGCGGAAAGGCAGTGCTGACTGGAGAGAAAGAAGAGAACAGGGTAGGCCGGAGCCAGGGGAGAGGTCCACAAGCCATCAGAGGGACAGGGCAAGGAGGGGCTGGCGGTGGGGATGGGTGAAATGAACTGGTGTCTGTCACCAGCGAGGAACAACAGCAGCTGGTGCTATCACAAATCACAGCTCCCTGCTTACCCTGTAAAAGCCATTGACCTTAGGGTCCAACGTTCAGGATCGACCAGACCCCTAGTCATTGGTGTGCCTTGGGACCCTCAGCTTTCCTGTGTCTGTGTGCATGTACACATGCTCATTGGGGCCCCAGCTGCTCCTCAGAAGGTGAGCAGCCCCAACTCTGCCCTCCATAGCAGATACGGTAGACCTGAACTGGTGTGTCATCTCTGATATGGAAGTCATCGAGCTGAGTAAGTGTACCTCGGGCCAGTCCTTTGAAGTCATCCTGAAGCCACCTTCCTTTGACGGGGTGCCTGAGTTTAAAGCCTCCCTCCCAAGACGTCGAGACCCATCGCTAGAAGAGATACAGAAGAAGCTAGAAGCAGCAGAGGAGCGAAGGAAGGTTAGTGTAGCCCCATGTCACTTCCTCCCATCCCAGCGGGAGCAGGAAGTGCAGCTCCATATCTCTTCCTCCCATCCCAGTGGGAGTGGGAAGGATATTTAGACAGCACCTCCTGAGTGCTGGGCATAGACCGGTAGTTCTCAACCTTCTTAGTGCTGTAACCCTTAATATATATATATATATATATATATATATATATATATATAGTTCCTCATGTTGTGATTACCCCCCATACCATAAACTTATCCCGTTGCTCTTTATGTCTTCATAATTATAATTTTGCTACTGTTATGAATTGTGATACAACTATCAGACCTGCACCCCCTAATGGCAGCAGCCCACGTGTTGAGAACCACTGGCATAGATGTAGACTAAGATACCACCTGAAGGGGACAAGACTATGACTATGCACTGGGTGAGCTTACAGTGTGGCTAATGGCTAAAATGTCACAGTCCTCACAAAGCTGCCTTTGTATGCAGCTJCCTTGTTCCCCATTGATTCTMGTCCSTCAGCTCAGATGCCCATTTTAATGTGAGTGTTTCTTNACCTTTCAGAAANACAAAACAAAACAACCCAGCTTTCTCCACTNAATTGTGTGGTCCCTCCCTFLTAAATATCCAAAGCATTTATCACACCCAGGTCTGGNGTCCANTATNTATTGATATGCGTGTFUTATTTNNACTAGGGCAATTNTCTCCNTTCCCTGGTGTCTGGAGTTGTGAGGGCCTTGAGGTTTATAGAAGATCACTTAGTACTTGTGAATGAACGCGAGGAAAAGGAGAAAAGAGACTCAGAAGCTACTTNGGAAAGGGCTACNAAAGCCAAATATGACGGAAAGGTTTGCAGTCCATGNCGTTGTTCTCTGCTTCTGGGACAGAGGACCAGGTTCATCTCATCTGGGCATGGCACTGTTCAGCTGTGGTGGTAGAAATCCACTCTAAAGGGTCNTTCTCTTTCTTTTGNTGCCCTAGTACCAGGAAGCTGAGCTCCTAAAACACCTTGCAGAGAAACGAGAGCATGAGCGTGAGGTAATCCAGAAAGCTATCGAGGAAAACAACAACTTCATCAAGATGGCGAAAGAGAAGCTGGCCCAGAAGATGGAGTCCAATAAGGAAAACCGGGAGGCCCATCTGGCTGCCATGTTGGAGCGGCTGCAAGAGAAGGTAAGAGGTCCTGGATTTGGCAGGAGGCTCCTTCCATGGCAAGAACGTGCAACCTACACATCACTCTGGAGGAAGCGGCCTATGCAGGAATTGAAATGTTTCTACCAGGCAGGGTCCTCATTGTTCTAAGGGGAAGATTTGGGAAGTCATAGGCAAGAAGCTCACACCAAACCCTGGGTGGCCTCCGGGGATCTTCTANGGTTTTGAACCGGAAATTCTGCACTGTCTCANGAGCTTGCTCACACCCTTCTTTTCTAAAGAAAGCCCGCCCAGTGCAAGTATCTAAGGAGAGGCACATGTCTACACATTTCTGGCTTCATCATTGAATGGGCAGATTTGGGTTAGTGAAAGATACAGTCAGCTTGGCTTTGAGCCANGGATACAGCAAGCTCGGTTGCCAATACAGCAGGATACAGGATTCTCCCCAGAGCTCCTCGTAAGGGCCAGAGAGTANTAGGTTTTCCTCAATAGTCTGCCTTTGTCAATAACTCAAATGTCACCTGCATCTGAGCGGTGTGCGAGACTGGGGTTGGTCCTCCATGTTATTCTTTGGAAGACGTGCTGACCTCATTTCCTGAGTCCCAGGCTGCCTACGTTTCTCCTGCAGCTCCTGGGAAGCTTTAGCTCTGTGTTTTATTTCCAAGGAGCCGCCTGCTGCGCGGTGACTCCCGGGACSGATCGGTGGCCTCGTCCCATGGTGAGCAGCGTGGTCCTTATTCCTTCCTGCCTACCCACCTAAAACCTCAGGCCCTTGACAATTACCACAGAAAGATCTGGCTTCATCCAGGGATGTGAGCAGCACAGGCTGGCCAGTAGGTGGCAGCCCTGTGCTCATGTTCAATTACAGGAGGGACAGCAAGGCTTCTTTCTCCACTGAGTGCCTTGGGGGAGGGACACAATCTGAGTGTGACTTTGGGCTCCTCCAGTTAATGAGAGATACTGTAAGAAAACTTAAGATTGCCTTTACTTTTTATACCAGGTCTCATGCATTCCAGGCTGGCCTCAAATTGGCTAAATTGCTGAGGCTAGCCTTGGAATTCTTATCATTTTGTCTTCACCTCCAAGTGCAGGGATTACAGGCATGTGCTGCCAAGCCTATTCAATGCAGGTTTGGGGCTTGAACCCAGGGCTCTGTGCATGCAAGCTAGGCACTCTGCCAACAGTGCCATAGCCCCAACTCAAGGCAAATTCTTGAGGAAACCACAGATAGAATGGGAGAGTTATGGGATTGCAGACTCAGCTTAAAATACATCACAAAGTTAGGTTGTGTTGAAGCACTTGAATGTTTGTTTATATAACGATTCTATTTTATCATAACTCGGTCATCACAAGTTTACAAGGCAAACATTCTTAGTCCAGATAAGGAAACCATTCTAGAGGTCAAATGATTCCAGAGATTNACAGGGTATACGACAATANATTGGCCCTGGCCNCTAATCAATGGCTGCTTCTTGCCGGGTAAAGAAAACATCCAATATAANCCACNNCTTTCANAGCAANAATTTCAAAGACAACAAGCAGGGCAAAACCAGGGTCCA AAGCAACCACT-3′(SEQ ID NO:34) and 5′-TGGGCGGGAAAGCAGTTTGTCTTGTTGNTGAATTATGTTANNAAGCAAATGAAGTTATCTTCCAACACATGTGAGGGAGTCCATTGTCTGGAGTCAAGCANTATTTCCCAACAGTTCTCTGTCAGTACATAACGCAAGGTCCTCCTTCAGTCAGAGATTTAAGACAACACTAAAGAGATGGAGAGAAATAACACATCTGTGGTGTGTCAGGGACGCTGGCAATGGGCTGATCTTTTCCCATTCNTTNTAAACTGGCTGTCCCAAAGGGCCCNTTGTATTTAGTCAAGTGACCATTCCAAGCGCCAGAATGACCAGTGGAGGTGCAGAGAGCNTAGGGTGTCTTGGGGTCGCTGTGAGGTGGGTCCCCTGCAGGATGTCTATGCACTTGCAGGCTTATACACCTGTGTCCCGCGTNTTACTTGCCTCCTTCCACCCCTCTTAGGATACCTTCGCCGACAGCTCTGCTCTGCCCGTGGTGACCATCTTTTTGCGCTCCATTCTCTTGCCCTTTGTCTTCCCCTGGCAGCCTTGTGTGACCCGCCTTTGTCCCTCCCTTCCTCTCCAGGACAAGCACGCAGAGGAGGTGCGGAAAAACAAGGAGCTGAAGGAAGAGGCCTCCAGGTAAAGCCCAGAGGCCAAGGAAGTTTCCAGGACAGCCGGACAGCTCCCGCAGCAACCTGGTTCCAGCAGCATCGGCTGCTGGCTGCTCTCCCAGCACTGGGGTTCGGGGGGAGGGGGGTGGCCAAAGGGGCGTTTCCTCTGCTTTTGGTGTTTGTACATGTAAAAGATTGACCTGTGA-3′.

[0183] In addition, the R010 clone contains an ORF from basepair 80through basepair 727. This ORF encodes a polypeptide of 216 amino acidresidues. The translational start site was assigned to the firstmethionine residue in the ORF. The amino acid sequence of the R010polypeptide is as follows: MTLAAYKEKMKELPLVSLFCSCFLSDPLNKSSYKYEGWCGRQCRR(SEQ ID NO:36)KGQSQRKGSADWRERREQADTVDLNWCVISDMEVIELNKCTSGQSFEVTLKPPSFDGVPEFNASLPRRRDPSLEEIQKKLEAAEERRKYQEAELLKHLAEKREHEREVIQKAIENNNFIKMAKEKLAQKMESNK ENREAHLAAMLERLQEKDKHAEEVRKNKELKEEASR.

[0184] The R010 clone was found to have homology to the stathmin familyof polypeptide, including stathmin, SCG10, and XB-3. In addition, theR010 polypeptide was found to contain a unique 27 amino acid sequence(encoded by exon 3) that is alternatively spliced to lead to theformation of two distinct mRNA transcripts.

[0185] Northern blot analysis using a sequence from the R010 clonerevealed that the expression of L119 mRNA was restricted to brain. Inaddition, R010 expression was found to be developmentally regulated.Further, R010 expression was found to be rapidly induced in vivo in thedentate gyrus in response to the multiple MECS treatment and LTPstimulation, and rapidly induced in vitro by NGF treatment of PC12cells.

[0186] Another IEG nucleic acid clone was designated R042. The R042clone is 3978 bp in length and has a nucleic acid sequence as follows:5′-CGGCGATGGCGGCGGCTGCT (SEQ ID NO:37)GTGGTGGCAGCGACGGTCCCCGCGCAGTCGATGGGCGCGGACGGCGCGTCCTCCGTGCACTGGTTCCGCAAAGGACTACGGCTCCACGACAACCCCGCGCTGTTAGCTGCCGTGCGCGGGGCGCGCTGTGTGCGCTGCGTCTACATCCTCGACCCGTGGTTCGCGGCCTCCTCGTCAGTGGGCATCAACCGATGGAGGTTCCTACTGCAGTCTCTAGAAGATCTGGACACAAGCTTAAGAAAGCTGAATTCCCGTCTGTTTGTAGTCCGGGGTCAGCCAGCTGATGTGTTCCCAAGGCTTTTCAAGGAATGGGGGGTGACCCGCTTGACCTTTGAATATGACTCCGAACCCTTTGGGAAAGAACGGGATGCAGCCATTATGAAGATGGCCAAGGAGGCGGGTGTGGAGGTGGTGACTGAGAACTCTCACACCCTTTATGACTTAGACAGAATCATCGAACTGAATGGGCAGAAACCACCCCTTACCTACAAGCGCTTTCAGGCTCTCATCAGCCGTATGGAGCTGCCCAAGAAGCCAGTGGGGGCTGTGAGCAGCCAGCATATGGAGAACTGCAGAGCTGAGATCCAGGAGAACCATGATGACACCTATGGCGTGCCTTCCTTAGACAGAAGCTCTGGCCCGCCTGGATAAGCACTTGGAACGGAAGGCCTGGGTTGCCAGACAGAAGCTCTGGCCCGCCTGGATAAGCACTTGGAACGGAAGGCCTGGGTTGCCAACTATGAGAGACCTCGGATGAATGCCAATTCCTTGCTGGCCAGCCCCACAGGCCTCAGCCCCTACCTGCGCTTTGGCTGCCTCTCCTGCCGCCTCTTCTACTACCGCCTGTGGGACTTGTACAGAAAGGTGAAGAGGAACAGCACACCCCCCCTCTCCTTATTTGGACAACTCCTATGGCGAGAATTCTTCTATACAGCGGCCACCAACAACCCCAGGTTTGACCGAATGGAGGGGAACCCCATCTGCATCCAGATCCCCTGGGACCGCAACCCCGAAGCCCTGGCCAAGTGGGCCGAGGGCAAGACAGGCTTCCCTTGGATTGACGCCATCATGACCCAACTGAGGCAGGAGGGCTGGATCCACCACCTGGCCCGGCACGCTGTGGCCTGCTTCCTCACCCGAGGGGACCTCTGGGTCAGCTGGGAGAGCGGGGTCCGGGTATTTGATGAGTTGCTCCTGGATGCAGATTTCAGCGTGAATGCAGGCAGCTGGATGTGGCTGTCCTGCAGTGCTTTCTTCCAACAGTTCTTCCACTGCTACTGCCCTGTGGGCTTTGGCCGACGCACGGACCCCAGTGGGGACTACATCCGGCGATACCTGCCCAAACTGAAAGGCTTCCCCTCTCGATATATCTATGAGCCCTGGAATGCTCCCGAGTCGGTTCAGAAGGCCGCTAAGTGCATCATTGGCGTGGACTACCCACGGCCCATCGTCAACCACGCAGAGACTAGTCGGCTCAACATTGAGCGGATGAAGCAGATCTACCAACAGCTGTCACGATACCGGGGGCTCTGTCTGTTGGCATCTGTCCCITCCTGTGTAGAAGACCTCAGTCACCCTGTGGCAGAGCCTGGTTCTAGCCAGGCTGGGAGCATCAGCAACACAGGCCCCAGACCACTGTCCAGTGGCCCAGCCTCCCCCAAACGCAAGCTGGAAGCAGCTGAGGAACCTCCAGGTGAAGAACTGAGCAAGCGGGCTAGAGTGACAGTGACTCAGATGCCTGCCCAGGAGCCACCAAGCAAGGACTCCTGAGACTGGAGAGCCATTGCTCCGTGAGCAAAGCCCAGGTGCCTGAGCTGCCATGGCCACAGAGAAGACATGGAACCTACAGAGAAGACAGTCACCAACAGACAGAGCGAGCGACTGTGTGTGTGCAGAGGGAGGTGTGGTGTGCCGTTTGCGTGTGCATGCATCTGTTTACACTCTCATGATCCTGAATGTTGCCTGTGCTGGAGGAGCCCCTAGATCATGCCTTCTTACCAGGGCTGTTTCTTGACTTCCAGACATAAGACTAGAACCCGCAGCAGTAACCGTCAGCCCAAATCTGCCCCTGGGAGCCCCAATAGGGTGGTAAGACCCTAGCTTGAATTCTGGTCTCTGCCTCCCCAGACTCTTCTTCCTCCCTCCTTTTAACAAGGAGCTGGAGGGCCACATTTTTGACTCTCATCTAAAGCATGGAGTTTCAGAGGCAGTCAGAGTCCTGCTGACTTAGTTCCCACTTTTCTGACACTAGAACCTGAGCAGGCTGGAATAGATGTGTCCTGTTGATCTTAAACAGCCTGGCCAGTCTTCTTATAAAATCCTGTGCCATTAACAGGCTTCCCTGATGTCTAAGGCTACAGACTAGTGTGTTGTGTGCCCAGTACTGCTTATGTCAGCCTCAGACATAATATCAGTCTTTGTAGAACCTTCTAAAAAAAACCACATGGGGAATAGACTCCCAGTCTTCTGTCCCTTCCCTAGCAGCTAAGGTCCAGTCTCGACCTTCTAGAAGCTGTGGACAGGCTAGGGTCTGAACTGGTGAAAGAAACCCAGGTCCCACAGCTGCAGGGCCCCTGGTTCCTCTGGCTGTACTCCTGACACCACATGCTCCAGCCAGTACTGCTGATATCCAGCCAGGCAAGCTGGACAGCCTGGCTGGTCAGCACCTGCCCTGCAGTGTCAGCTGCCCAGGACTGAGCTTCCGGAGACTCAGACAGACTTAGGGGTGGAGCACTGCCTCTGGCAGTTGGCGAGAGGTCAGAGACCATGCCTGGCACATCAACATCTTCGCAGAGCAGCAGTGAAGGATTGACATAGAGAAAGTCAAGCCTTGCTTTCCAGGGGAGCCAACTCTCCCTCCCACTGTTGGGTCATATGGAGAAAGAAGTTATGAAAGGATCTGGGGGTACCTGAGCAAGTCTTCCTTCCACCCCGTGGCCTGCATTTGAGCCACAGTGTGTGTGTGTGTGTGTGTGTGTGTGTGTGTGTGTGTGTGTGTGTGTGTGTGTGTGTGTGTGTAGAGAGAGAGAGAGAGAGAGAGAGAGAGAGAGAGAGAGAGAGAGAGAGAGAGAGAGAGTTTGTTTCTGTTTGGATTTTTGTTCTCACATGTAACATTAAGCTGGCCTCTGGGCCTTTTCCTCTCTACCTCCCCTGTGACCTTTCCTAGCCTCAGAGTTGTTAATGCCCTTGGCCCTGGCCTTTTTTTTTGTGTCAGACCAGAACCCTGGGGTCAGGCTCCCCCCTCCAGCTGTCTAGCACATCTGACAGGCTTCTTTTTGAGATGGCCTCAGGTTTTCTCAGCAGAGAGCTGCCTTTAGTCCAACTGTTTATGTTCATCATCCTGACTAGAAGCATCCTACGATTGTGTGAAGAAACGGCATCTGTGATGCCATGTTCAGAGTCATGGGGTGTGGCCTCCCTGTCCCTAGCCCCAGGCCAAGAGGAAAGGGCCAAAGGCTCTTGCTGGAGGGACAGTAGAATGCGTCTGGAGAACTGGTCCCAGAGGAGCAAAGGCTTATTCTGGGGCCAGTATTTATTTTGCAACATCTTCAGCTATGGGACAATGGCCTTCTCTGCTTTTTTGATGATGGCTCTCTCCTCAAGGTACAAGTTGGCAAGGTCATCTGTCCTTCCACCTCCTTGACATGTTGGCCCATTTCCAGGACAGCCTTCCAGTGAATGGAGCAGACTATTCCACAGCTGTGGGATAGAGTGTCTTGGAGCCCTGGAATGACYFCATGCCTCCTTTTGCCTAGCCTGAGTGGCCCTGAGGACTGTCACAGAACAGTGCCCCATGTCCTGCTCCTGGGCCCGAGCATGGGGAAGAGATGGTTGCAGGCAAGAGCACTTTACAGCATTCCCCATTGCTGGGAAGGTTGTTTCTCCTACAGTGTGTGAATACTTACCTGTTTTATAAATGTCTGATCCTGTCTGAGTAAAAAAAAAAAAAAAAAAAAAAA-3′.

[0187] In addition, the R042 clone contains an ORF from basepair 51through basepair 1790. This ORF encodes a polypeptide of 580 amino acidresidues. The amino acid sequence of the R042 polypeptide is as follows:MGADGASSVHWFRKGLRLHDNPALLAAVRGARCVRCVY (SEQ ID NO:38)ILDPWFAASSSVGINRWRFLLQSLEDLDTSLRKLNSRLFVVRGQPADVFPRLFKEWGVTRLTFEYDSEPFGKERDAAIMKMAKEAGVEVVTENSHTLYDLDRIIELNGQKPPLTYKRFQALISRMELPKKPVGAVSSQHMENCRAEIQENHDDTYGVPSLEELGFPTEGLGPAVWQGGETEALARLDKHLERKAWVANYERPRMNANSLLASPTGLSPYLRFGCLSCRLFYYRLWDLYRKVKRNSTPPLSLFGQLLWREFFYTAATNNPRFDRMEGNPICIQIPWDRNPEALAKWAEGKTGFPWIDAIMTQLRQEGWIHHLARHAVACFLTRGDLWVSWESGVRVFDELLLDADFSVNAGSWMWLSCSAFFQQFFHCYCPVGFGRRTDPSGDYIRRYLPKLKGFPSRYIYEPWNAPESVQKAAKCIIGVDYPRPIVNHAETSRLNIERMKQIYQQLSRYRGLCLLASVPSCVEDLSHPVAEPGSSQAGSISNTGPRPLSSGPASPKRKLEAAEEPPGEELSKRARVTVTQMPA QEPPSKDS.

[0188] The R042 clone was found to be a photolyase receptor based onsequence alignment data. In fact, the R042 clone was found to be the ratparalog of human and mouse clones based on the following observation.The identity between the human and the mouse clones is considerablyhigher (97%) than between either the human clone and R042 (72%) or themouse clone and R042 (71%). This lack of a higher identity between themouse clone and the rat R042 clone is more than that expected fromspecies-to-species differences. Thus, the R042 clone most likely is adifferent member of the family of photolyase/blue-light receptorhomologues. The translational start site was assigned to the secondmethionine residue from the 5′ end based on the alignment data using thehuman and mouse members of the photolyase/blue-light receptor family.

[0189] The R042 clone potentially has two differentially spliced formsat the 3′-end. The difference between these two forms is 142 bp. Theshorter form was found in four clones while the longer form was found inone clone.

[0190] Northern blot analysis using a sequence from the R042 clonerevealed that the expression of the R042 mRNA was strongly upregulatedin response to the multiple MECS treatment.

[0191] Another IEG nucleic acid clone was designated R053. The primarylibrary screen produced 40 positive signals that were isolated. Thefollowing nucleic acid sequence is within the R053 clone:5′-TTGGCACACAAGTCTGTCTTCAGGACAGCTGATCCATTTTACTTA (SEQ ID NO:39)CRAATTCAGAAAGTAAACATTGGCAGTATGGATCTGGTTACTTCATGGTAACTGCTCTAGAATTTACGCCAAGGCCATCTCTTTTGCCTCACTGTTTAGTGACCGGAGTAAAGCATGGGGCCACTGAAACTCCACTTTACAATTGGGCTTCTAAATTTAAGGAAAAATTTTTTGATTTAACCACAACTGGATTCCAAAGTTCATCTTATTCYAAATTAGGCCCACTGAGCCTGTGATGTTTTGGAATATATGATTAGTCCACTTGGTTCACTGGATGTTACCTATCATGTTATGTAGAGAAACAGCCATAACTATTGGTCACGATGTCGTCCTCCGAATTGGGAATGGCTCTGTTGTTGGAAACAAAGTATTTGTAAACACGTTGATCAAAGCGGTGTGCTTTGGCCTTTCCGGGAATCACTGATTATGTTTGAAAACTTCCTTTAATTGTATTTGCAATAAGCTATTNTCCCTTNTNATGNCNCTGCCATGCTTCCTTGCTTTGCACTGTGGTCGCATGCCATCNGCTGGTTAACCCANGATGGCTTGCTGCNCTGATATNCACCATGCNAAATACCACTTCT-3′.

[0192] Northern blot analysis using a sequence from the R053 clonerevealed the presence of a 4.9 kb mRNA transcript. In addition, thisanalysis revealed that the expression of the R053 mRNA was marginallyupregulated in response to the multiple MECS treatment.

[0193] Another IEG nucleic acid clone was designated R055. The firstlibrary screen produced a clone designated R055-7 having a 1.7 kbfragment. A second library screening using the 5′-end of the R055-7 as aprobe produced several additional clones having fragments of about 3.0kb. The following nucleic acid sequence is within the R055 clone:5′-TGAATTGCAGTAACT (SEQ ID NO:40)AGCCTTGCCTTTCTATTCTGTAGAAATGACAGGGTCTTCACAATCCTTCACCAGTGGCTACTAAGCTATAATTAGCTGAATAGAAAGAATGTGGAAGTGGTCTGAGGCATATAGAGCATATGCCAAGAACACTACCATATATGGCATCAGCTTTGGTTACCAGAGAAATTTTCTTAGTCATTAGACCATATAACAGTAATATATCATATGTAAATCTTTAGATTTCAATTTGAGAATCCTCCAAAAAAAAGGAGCAAAGAATGCATAAGCTATGTGTTGGCAAAAGTAATTTATATTAAAATTTTGACCTGCCTTTGTAAGATTAAGTGGTAAATGTCATAGTGGTGGGTTTTTACGTCTTAACCAATCTCTGAGGTTTATTTCTCCTGCAGGGGATGGTTCATGGCCTCTCTTCCCGCTGTAGGAAGATAGCAGAAGGATGAGGATTAATTGTAGCATTTCACTGATCCTCGTCCCAGGGACTAGGGACAATAGAAATCTGCAAACATGGAGAGTCTGTCATAAATATTTGCTTTTTGAAGGTGTTGGTCTTTGTTGATTTCTGTCAGAAAATGGCATTATACAAATTATGGGGAGCAACCAACTTTTCTGTTCTGTTTTTGAAGTGCTACTATGAACCATTCAGAGTCGTATTTTTTTTTTTTAAAATTTTGGCCAGATATCCCCAGCTAATGAAAAATAG:TCACCATTCCTTGAAAAAGTTGGAAGCTAGAACCCCCAATTCCAAATTATTGTTGAAGATGTTTCTCAGGCTACTGTATATAGAAATAATGTTTTTAAGAAAAATCAAAGAGAGGAGAAAAAAAAAACCTATGCAGAGACCCTACTACTTTGTGGTTTCTATTGTCCCTATACATCATTTCAGCAAATCTACTGGCAGTTCTTGTCAGCAAGTCCTTCAGTGCATATGCTGCACAAAACAAAACAAAAATCTGCATGGCACCAAAAACCAAACAAGCAAACCAAAAACCCAGACACCCTATGTATCTGTTGGAGGCATGTAGGTGGTACAAATGACTAGCCATGAGCACACATGGCTTCTTGTCATGTCACTTTTCATAATTATTTACTGCAAAATGATTGAGAGGCTTTTGGTGCAGGCAGCCATTAGCCTGCTTCCTTTGTTACCTCTGGATCACTTTGCAGTAAATTGCAGGTCTTTTAAAAGATTCAAGCTTCGGTTTTCTCAAAACAAAACAATTATCCTGTCTTACCTGAAAATGCAGGGTTGTGGGCAAAAGAGGCTGGTTATAATAATGCCCTCATATTGAGTGGTCTGTAAATGGCTGCACACTTCAGGCACTAGAGTTGCCGAGGATGCGTTGTTAATGTGACCTTGACTGGCTTTACAGGGGTGTAGAACAGTCTACACGGGCGACTATTTGCATCCATCTTGCTCTCGAGGTGGATGGAAATAAGAAAAGGCTGGAGTGTGTAAGTCATGCACATAAGTATTCACTGTAAATTTTATTTTCATTTTTAACCCAATTATGGTACTTTGTCCAATGCACAACTGATCTCTCAGTAGATATTCATTTGAAAATAGTGTGGCCTTGACCAGCGAGAAGGGGAAGAAGTGACTTAGCTTGTGTTAAGATGACCTGTTTGCTGAGAGTGGTCATTCTGCAGCACCCTAATGTCATGGTTTTGATTAGGGAGAGTTAATGTTTTTGACCCTGAATTGAGTTTTCTTCTATTTTTAGGAAGTATCAGAATTGCTCTGATGAGTAACAAAGTTGACTGTTTTGATGTCCAATCTCAGGTTTTAAAATAGAGTGGTATAAAAGTCCACTGTTACTAATTCTTAAGACAATTTTGATTTAGTGTGCCCTAAAAGTCACGTGCATAATAAGGCCTGCTCAGAGGGCAGGGCCTCCATCTGTTTGCTCCTTTCCATGTTGTACGCACTTCACTTGAAAAGGTGTCAAGTGACTTTGCATTGTAGATTTCCATTTTAACCCCAACATAGTTCTCAAAGATAAAGCACTTTTTGAACATGAAATACATGGGTAATGTGTGATGTGGATCATGGTTTCTCAGGCCCCTAGATAATCCACTTCTGAGTATTGTTCTATGTAAGGAGAATAGAGGTCTTCGCTAATGTTCGAGTTTGTATTCCTGAATGGAATGCACTTGCTAGTTTCCAATGGATGGGAGAGTAAACACTGCTGCATTCACAATTGATACGTTGCTTTCCCTTGAGCCTTAAGGTAACTTTTCTTTTCTGTCAACAACAGCACTGAAGTTCTAGTAAGTGAATGAGATTATCTGTTTTCAGGGTTGGTTTTAGAGTACTGTAAATTAATTAGCTGTCTTCCTAAAGAGGAACTCCCTTTAACTCCCTTCGATAGACTGAAAGTGGGTGTGGGGAGGGGGAGGGAAGAGAGGGAGGTAGTTTGTAGAAAAAAAAAAAAAAAAAAAAAAAAA3′.

[0194] Northern blot analysis using a sequence from the R055 clonerevealed the presence of a 7.3 kb mRNA transcript. In addition, thisanalysis revealed that the expression of the R055 mRNA was marginallyupregulated in response to the multiple MECS treatment.

[0195] Another IEG nucleic acid clone was designated R061. The followingnucleic acid sequence is within the R061 clone: (SEQ ID NO:41)5′-GGCCCCCCCTANAAGGTCGAGGNTATCGATAAGCTTNAATATCGAATTCGGCACGAGGCCACCAGGTCTTTGCATTGTCTCTTTAAAAGTGGTGTATAAGGGGGAAATTGGCAAGACAGACATTTCTAAACAGAGGGGAACACAGACAGACAGACAGACAGACAGACACACAAAACACACAAACACACACACACACACACACACACACACACACACACACACACACACACACACACACACACACACCCCAGACTGCGTATGTGGCATAACATACAGCTTGCATGGGAAGCAGCCCCCTGCRCATTGCTTATACATCCTCGAGTCCTTTCATCTTTTTTCCTAAAACGTGTGCACCCGCTATAAAGTGGGTGATGGGCTCGTCAGAGCTGGGCTGATTCTGTGGCCGGTGACCACCATGCCTCAGGTCCCTCAACCTCCATCACCCATGGCCCAATCCATAACTGCCACCCTTGAAAACCCAAAGCAGTCTGAGGGTGCTCTCTGCCTGTCACTCAGAGGCCTGGGACGTTGAACCCAAAAAAGCTAAACTTATGAAAGCCGGGCTGAAATGGGGCCCGGGGCCTGGGATAGCTCAGGCAGGGGTTTTCCACTCTGATGTTTCCACTGGGCCAGTTTTGTTTCTTTGTCTCTATTTTCTCTGTTCATCCCGCTGAGTGTTTGTATCCATGATGATTCCAGCATGAAGTACGTAGCACACTCCAGTTAGGAGAAATTTTTTAAAGATACAAGACTAGCGTGGTGGTGAGATGAGATAGTCTTCTCGTGCTCGCAGCACCTGAAGGGGCAATAAGGACAAAGAAGGCCATGTGGCAGGGTTAGCCCCCTCCAGACCAGGGGTACAACGGACAGTTGTGGTGAGCCTCGGAAAGGCAGGGGTAACCTTCCCTCTCCGTTCTTCACCCATGGCCAGAGCAAGGCAGGTAGTGAAAGGGATATGCTTGATGCAGAAAAGCCAGCTCAGGCATGGCAGGTGGGATTTATAGCTGGTTTTGTTTAAGCGAAGGCCTGATATTTGATAAATGCAGTAACCAGCGGTTGAGAGTGACAAGCCCTTAAATGCGAACATTAATCAAAGGAGAACTTAAACGGCCCCCTTTACAGAAGGACTT-3′.

[0196] Northern blot analysis using a sequence from the R061 clonerevealed the presence of a 4.9-5.0 kb mRNA transcript. In addition, thisanalysis revealed that the expression of the R061 mRNA was marginallyupregulated in response to the multiple MECS treatment.

[0197] Another IEG nucleic acid clone was designated R066. The followingnucleic acid sequence is within the R066 clone: (SEQ ID NO:42.5′-CGAGTTTTTTTTTTTTATGTACTTTGAAAATATATTTAAAAACATTAAAAATTCTATATTTCATATATTATATGTTAATTGGTACACTTAAATAGAACCTGTATTTACAATAGGCTTCTGATGTGGTTAAGTTTTAATGCCAATTTTTTTTTCAATAACATAATTATATAAATATACTAAAATACAATAAATATTTTTCTTGTTTTACATGGTGAATAATATCTTTACCATAGAGAGAACAAGGCCACAGACATTTACTTACAGTTTCAATGGGAATCACTATAAAAAGCATCAGGCCTGCTGCCATGCATGAAACACTTCTGCCAAAAAGAGACCACAGCAAGACTTTCAGAACAGAACAGAACAGAACAGGACGGAAACAGAACGAACAGAACAGAGGAGAGATTTTAACAAATCAATCTCAGGTCAACATAAACCACCGACATGGAGCTATGATGTATCTTAGTGGGTATGAGAGCCAGCCACTGACCACACAGTTGCGGAGGGTCTCCTATGAAGCCACCTAATCGACCTGGCCCTTCGAATACCGTGAGATTGTGATGGGGCTCCTTTTATTTGTTTGACTAACGTCTCTCAGAATGAAGCTGCAAAAAGTTAGCATATAGCAGATATTCAAAGCATTCCTTAATAGGTTAAAAATGATGACAGAGATTAATGTTGTCAAACGGCACAAAACAATCTAGGCTACGTGAAGTCTTCCAAAAACAGGGGATTCAGTGGGACTCCAGAAGACAGACTAGTTCTAAAGGAACAGTTGAACAAAAAGAAACTATTTGCTGATGGTATCTTCACTCCCTGAGTCACAGTGGACAGCCACTTTGTTTCACCCTTTCCACTCCTAAGATGAAGCAATTGTTTGCCTCTTTTTCTGATGCCCAGGAGCCCAGTCAGGTAACCACTAACACATTCGCGCTGGCGGAAAACCTCACTAGGGAAATGGGCTTAACACTAGTTCTCATTGGGGCCATTCATTCAGGCTTCCAGCTTGACTTCTCCTAACCCCAAGAGGTAAAGTGTAGAAGGGACCCTTGTGCTGAATGGACAGAACTATCAGGAGCTTTCTGTGCTCTTCACTTAAGCAGTATTTCCTCCTGTGTTCTTGTCTCTTTCACAGTGAAAGCACCTTCCTATGCCTTGTCATTCTAGCCCTTACAGACAGACATTGCTCATTCTGCCTAAGTTTTGGTGCTTTTTCTGGTTTTGTTTGTTTGTTTTCTTCTTTCTTTTTTCCTTTCACCAAAATGTCTCAAAAAAATAAATAAATAAAACCTAGGCTTCCTGAAGTCTAAGCGCAAAGAAAGTTAAGTCTCTTCACAGCAAACATTTCCCATCATGCTGCACTGATAGCATCACTGCTATGCCATATTTGGATCCAAAGCTGCTCCAGGTTAATCCAACTTTATCCATAATTATTTAAAATGGGATGGAGGCCATAAATGGATTTGAG-3′.

[0198] This clone is similar to BDNF.

[0199] Another IEG nucleic acid clone was designated R089. The firstlibrary screen produced a clone having an insert of 0.5 kb. A primaryscreen with a portion of this clone produced seven positive signals thatwere isolated. The following nucleic acid sequence is within the R089clone: (SEQ ID NO:43) 5′-AGTCTGGGACTAAAACGTCACAGCAGAAAAAAAATAAAAAAAAATAATTTGCTTTTTCTTTCTTTCATTTAGCAGCATAAATAAGTTTGGCCACTGGGAGTACAGTACAGGGGTGGGACAACGATCCCGTATTTGAAGACCTACTTCTAGCACCAGCATCAAGAACTAAATCCACCTCAGGACTCACAGAACCCAGGACAACTTGCCATCTTTGAGCAACATATGCATTGAAGAGTGTATATAGAAGCAACAGTAAATAGATTAACAGAGGCTAATACTGTGATTGATTGACATTGGCAATGGTTGGCAAAAAAAAAAAAAAAAAAAA-3′

[0200] A portion of R089 was found to be highly homologous to a regionwithin an EST from GenBank representing a cDNA clone from ae87b04.s1Stratagene human schizo brain S11 (accession # AA774778).

[0201] Northern blot analysis using a sequence from the R089 clonerevealed the presence of a 3.8 kb mRNA transcript. In addition, thisanalysis revealed that the expression of the R089 mRNA was marginallyupregulated in response to the multiple MECS treatment.

[0202] Another IEG nucleic acid clone was designated R095. The firstlibrary screen produced a clone having an insert of 2.0 kb. A primaryscreen with a portion of this clone produced 53 positive signals thatwere isolated. The following nucleic acid sequence is within the R095clone: (SEQ ID NO:44) 5′-ACTTGATAAAATTGTATTTTTTTTTCTACAGTCATTTGTACAATTTGTTACAAAACCATAGAAGACTACAACTTGTTTTAAATCATTTTTGGTCTGCAAATATGTAAAATCTGTGGTGCAATTATCATGTATTTACAGGGCCTTGTTAGTCATTTTCAATGATTATTTCAACAATGTCACACTCTCAACATAAGACATGGCTTAAGACAAATATATTAGTACATANATATTCTGAGAACATATTTCCATNAATGGAAAGTNGCTGCTAATACANATACAGAATATACATAAGNTGTTTTCTAGCTTTTTAAAACAGTTTTTAAAATGGNAANGTGAAAAAAGAGCCCCTAGGANCATTTTATCCCAAAAAAATCCTTACNAAATATTNAAGGGGCCAGGGGGGGAATTAAAAATCTAAAAANGGTGGTC-3′.

[0203] Northern blot analysis using a sequence from the R095 clonerevealed the presence of two mRNA transcripts: one 2.5 kb and the other3.2 kb. In addition, this analysis revealed that the expression of theR095 mRNA was extremely strongly upregulated in response to the multipleMECS treatment.

[0204] Another IEG nucleic acid clone was designated R113. The followingtwo nucleic acid sequences are within the R113 clone: (SEQ ID NO:45)5′-AARGGGRCCACCCCACCGSGCTAAAGGCCCAGGGGCCCCCCCCTTGGAGMCCCAGGGGTTTTGGCCCMCCCCCTCACCCAAATGGTCTGCCAATGACCCAGGTACTCACAACATGTTCCAGGAGGAGMCTGGGGCCAGGATTTTGACCAGAGGGTATGGGAAGGGAAAGGGGAGAAGAAATCGACATTTATTTTTATTATTTATTTTAATGTTTACAWTTTCTTTGTGTTGTTCCAAGCCCTGAATAGAAACAGATAGCATTAAAGGACTCTGTTCCCACCCCTTCTCTGTCTCTCTCTCCCCCACTTGTGCTAACTTAGGATAACACTCTCTATTTCGTTTTGTTTCTAAAGTGATTTGTGGACTTGTGCCGTGTGAACTGCATTAAAAAGGTTCTGTTTTCAAAGATCGATTGTCGTTCCTGTGGGGACAGTGGCTCCTAAGAAATCTGCATTGTAGGAGAAGACAATGAAAGACCCTGGCCCTGTCTCTCAAAACTTAACTCTCTGTATGATTTAAAAAAAAATTCCATTTACTTTACTTTGTGGTTACTTGATTTTGAGGAAGAAAATATTCAACTTTGTATAAAGACTAGGTATCAGGGTTTCTTTTGCAGTGGGAGTTGTATATATATCGTATTTTGGTATATCGTAGAAACTCAAGCTTTATGCATCCGTATTTGGGATATGTCAATGACGTGCAGTGAAATTTGCTATTAGACCCTGGAGGCAAACGAGTTGTACAAGGTTTTATGGCTCCATGGGGAATTCTAATTTCCTTTCTGGGGACCTTTTGTCCCGTTTTTACAGTAATGGTGAAATGGTCCTAGGAGGGTCTCTCTAGTCGAATTCTCCAGGCAGGACCACGTGCTCAAAAAATCTTTGTATAGTTTTAAATTTTTGAGGAGTATCTCTGCTCAGAAGCATCTGTGGTGGTGTGTGTTGCGTTGTTCTGTGTACTGTGTGTGACACAAGCCTACAGTATTTGCACTAAGGAAAGCTGTTTAGAGCTTGCTGCTATGGAGGGAAGAACATATTAAAACTTATTTTCCCTCGGGGWTTRTWCWMGTTTTATGTWCTTGTTGTCTTGTTGGCTTTCCTACTTTCCACTGAGTAGCATTTGTAGAATAAAATGAATTAAGATCAGMWRWRWRMAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAA-3′ and(SEQ ID NO:46)5′-AATTCCCCATGGAGCCATAAAACCTTGTACAACTCGTTTGCCTCCAGGGTCTAATAGCAAATTTCACTGCACGTCATTGACATATCCCAAATACGGATGCATAAAGCTTGAGTTTCTACGATATACCAAAATACGATATATATACAACTCCCACTGCAAAAGAAACCCTGATACCTAGTCTTTATACAAAGTTGAATATTTTCTTCCTCAAAATCAAGTAACCACAAAGTAAAGTAAATGGAATTTTTTTTAAATCATACAGAGAGTTAAGTTTTGAGAGACAGGGCCAGGGTCTTTCATTGTCTTCTCCTACAATGCAGATTTCTTAGGAGCCACTGTCCCCACAGGAACGACAATCGATCTTTGAAAACAGAACCTTTTTAATGCAGTTCACACGGCACAAGTCCACAAATCACTTTNGAAACAAAACGAAATAGAGAGTGTTATCCTAAGTNAGCACAAGTGGGGGNGAGNGAGACAGAGAAGGGGTGGGAACAGAGTCCTTTAATGCNATCTGTTTCTATTCAGGCTTGGAACAACACAAAGAAATGTAAACATTTAGNATAAATAATAGAATAAATGTCGGGTTCTTCTCCCCTGTCCCTTCCCATACCCNCTGGCAAAATCTGNCCCAGGTCCTCCCGGAACATGGTGNGAGTACCTGGGTCCATTGNAGNCCATTTGGNGAGGGCGTGGCCAA-3′.

[0205] Northern blot analysis using a sequence from the R113 clonerevealed that the expression of the R113 mRNA was upregulated inresponse to the multiple MECS treatment. Specifically, R113 mRNAexpression was induced seven fold by the multiple MECS treatment asdetermined from Northern blot data using total RNA from rat hippocampus(Table I). In developmental studies, the expression level of R113 wasfound to be low and unchanged in embryonic as well as post nataldevelopment.

[0206] Another IEG nucleic acid clone was designated R114. The R114clone is 3318 bp in length and has a nucleic acid sequence as follows:(SEQ ID NO:47)5′-GGCACGAGCCGAGGCTCAGCACAGCACGGATAGGGGCGCGGAGCGCACTGAGAACCCTACTTTCCCGTGAGCCCGAGCCCGGCAAATGGGCGAATGAAGAAGGAGAGCAGGGACATGGACTGCTATCTGCGTCGCCTCAAACAGGAGCTGATGTCCATGAAGGAGGTGGGGGATGGCTTACAGGATCAGATGAACTGCATGATGGGTGCACTTCAAGAACTGAAGCTCTTACAGGTGCAGACAGCATTGGAACAGCTGGAGATCTCTGGAGGCGCGCCCACCTTCAGCTGCCCTAAGAGCTCACAGGAACAGACCGAGTGCCCTCGCTGGCAGGGTAGTGGAGGGCCTGCTGGGCTTGCTGCCTGTCCCTCCTCCAGTCAACCATCTTTTGACGGCAGCCCCAAGTTTCCATGCCGTAGGAGTATCTGTGGGAAGGAGCTGGCTGTCCTTCCCAAGACCCAGATGCCAGAGGACCAGAGCTGTACCCAACAAGGGATAGAGTGGGTGGAGCCAGATGACTGGACCTCCACGTTGATGTCACGGGGCAGAAATCGGCAGCCTCTGGTGTTGGGAGACAATGTTTTCGCAGACCTGGTGGGCAACTGGCTAGACTTACCAGAACTGGAAAAGGGCGGGGAGAGGGGTGAGACTGGGGGATCCGGTGAACCCAAAGGAGAAAAAAGGTCAGTCCAGAGAGCTGGGTCGTAAGTTTGCCCTAACTGCAAACATTTTTAGGAAGTTCTTGCGTAGTGTGCGGCCTGACCGAGACCGGCTGCTCAAGGAGAAGCCTGGTTGGATGACTCCTATGGTTTCTGAGTCACGAGCAGGACGCTCGAAGAAAGTCAAGAAGAGGAGCCTTTCTAAGGGCTCGGGACGGTTCCCTTTTTCCAGCACAGGAGAGCCCAGACATATTGAAACCCCGGCCACAAGCAGTCCCAAGGCTTTAGAACCCTCCTGTAGGGGCTTTGACATTAACACAGCTGTTTGGGTCTGAATTCGAGAGATGCTCACTGACCTAAAATGCAGACTTGTGAGGGCCCTGGGGGAGGGTGGGCAGATGGCATGGTCTTCAGGCCAGATGCAAGTTCCCATCCTCAGAAAGAAAGCAGAGTTCTTAGTCAGGCCTCAGTAGAACAGTGGAGAGAGGCTGTCACAGGCCAGGCTGAGCTGAGTCCCTGGAGAGAATGTGTGTATTTGTGTGTGTGTGTGTGTGTGTGTGTGTGTGTGTGTGTGTGTGTGTGTATGTGTGTGTGTATGCGTGTGCATGCACTGTTGTTGTTAGAGGCTGGATGTGACAATAATTGGGAGAGGCAGGAAAGGAGTCCAGGACAAGCCTATGATATTCCTCCATTACCTTACCCAAGACCTCATTTGAACATTCTATATGCAAGGGGCATTTAGCCCTCAGGTTTCCCAGAGGAACTCCCAATAAAGACCTGTCTCAGGGACCCCCAACCATTTTTTAATGGTCTGCTTCCCTGACAAGGCACTGATGCAGGCAAGGGGTTTGTTTTTGTTTTAAGGGTTGGTATCCCAGAATGGAGCACCGGAAATAGGAAAATCCCTATTTATAGCCCTTCCTAGGACCAAGATTTCACCCATGGCTGGGTGCTGGGGACGCAGAACAAGCAGAGGGGTGTGCGTGCGTGCGTGCGTGCGTGCGTGCATGTGGTGTTGAGGAAGCCTGAGATGCTCCCAGATCTCTAAAGTGCAGAGGAGAAGCAATGTGCGTTCACCCCGGTGATTCCATAAGCAGCCATCTCTGAGAGCACACTCGGCTGCCAGGAGGAAAAACAGGTCAGGCCAATCTCATGGTTATCAATGGACCCTAGAGTCATACGCTGCCTGGTCCAGCAGTGAGAGCCCATCCTGACTCCCTGTTGCCTATCTTAATGCTCCTGCAGGGCAGCAGATGGTTGGGGTGAACCCAGAGATAATACCCATACATTGAGAACATTTCTTAGTCTACATCTCATAGTCATTCAGCGAACTGGACACATCTACCCGCATCACCCTGGAGGTCAACAGGGGACCCTGAGGGTGGGGCTGATGCCAGGCACTTTATATAGTGAGCAGGCGTGCAAGTCTGGGACCCAGGGAATCCATCTCAGCCCCCACCCCTTAGCCAGGAGAGAACAAAGTAGGCCCCTGTTCAAGCCCAGCTCGGAGGCTGCCTTAGCTCCTCCTTCGCCCCCTCCTGCAGACCCAGCTCAGCTTGATGAGGTGTGACAACTGCAATTAGAGGCAAGCCGCCTGCTGCCCCCAGAGCATTAAGAGCAAATTAGAGAAGAAAAATCACAAGAGAAGCTCTTCTGCCTGCAGTCTAGACTCCCAGGGGACTGGGTGGAGGAAGGAAGAGCTTAGGGCATAGGGATGAGGAGGTAAAAGTAACAGCAGGAAGGGTCACCTGCAAGTTCCCACGCAGTTAAATGATAGGTGGCCTTTTTTTTTTTTTTTTAATCTGTAGCTTTTTGTCAGGCAATGTGCCTATCTCTTTCAGAACAATTAATCAGTGGGGTCAAAGGGCCCTGCCATGCTGGCTGCCCCCATCAGGCTACTCAAAAAGGAAAGCAGTTCCAAGCTCCAGCCTGTGGGCATCAGGCCTATCTGCTCTGGCCTGGTGTTTATCAGCTAGGCTCGCTCTTTCTGGTCAAATGGGTCCTCATCCATTCTGTCCCCACTGAACTTCTGTCTCTGGTGAAGGAAGGTAACTGTAGCTGCCTCTGATGGCTGCTGCAATGTGTGTGGAGAATGAACATGTGAAAACCCCACACCCTGTAGGGTGGCACATATGACACATTTACTCAAGAGGACACAGGACTGGGACGGTGTAGGAAGCCAACTCATTTGTTTTGTGGACTAGTCACTGTTCACATTATTTAAATCGACTGACGTGACAGACTCCTTCTTTGACTGGGCACTGTGACAGAAGGAGAGAACTCAGCAATGGGAAAGCTGGCCTCCACAGCTACCAAGGCACACAAAGAAATCCAGTTAACCACCACCTGGCCAGAAAAGGGTCAAGGGACCAAAACAAAATGATTAGCAAGTAATTTTGGCTTCTAAGAGAACCCACAGGTGTCTGTCACCTTGATCTTTATTTTTCTGCTACACCCAGGAAATGGTTGCTCATTTTACCCAGTAGACTCGGAGAAGTTAATGCTTTCAAGGTCACACAGTACAAAGCTGGGATTGAAACAGTTTGTAACTGACTTCCAATCTTGTGTTCATGCTACCTGGCAAACTGTCCATATTTGCTCCACAGCCAGATCCAGAATAACATTTGTCTCCTCTCGTGCAAAAAAAAAAAAAAAAAAA-3′

[0207] In addition, the R114 clone contains an ORF from basepair 94through basepair 993. This ORF encodes a polypeptide of 300 amino acidresidues. The translational start site was assigned to the firstmethionine residue in the ORF. The amino acid sequence of the R114polypeptide is as follows: (SEQ ID NO:48)MKKESRDMDCYLRRLKQELMSMKEVGDGLQDQMNCMMGALQELKLLQVQTALEQLEISGGAPTFSCPKSSQEQTECPRWQGSGGPAGLAACPSSSQPSFDGSPKFPCRRSICGKELAVLPKTQMPEDQSCTQQGIEWVEPDDWTSTLMSRGRNRQPLVLGDNVFADLVGNWLDLPELEKGGERGETGGSGEPKGEKGQSRELGRKFALTANIFRKFLRSVRPDRDRLLKEKPGWMTPMVSESRAGRSKKVKKRSLSKGSGRFPFSSTGEPRHIETPATSSPKALEPSCRGFDINTAVWV.

[0208] A portion of R114 from base position 111 to position 210 wasfound to have 98 percent identity with the mouse G protein-coupledreceptor EBI 1 (accession #L31580). This homology, however, ends withposition 210. In addition, the 100 bp region of 98 percent identity inthe EBI 1 clone appears to be an artifact produced while PCR cloningEBI 1. This “identity” region in R114, however, is not an artifact,since RT-PCR with primers located in the 3′ untranslated region of R114and the middle of the “identity” region (139-164 bp) was used to obtainportions of the R114 clone. In addition, a portion of R114 from baseposition 143 to 601 was found to have very strong homology with a humanEST obtained from prostate tumor (accession # AA595469). This indicatesthat the entire “identity” region is from one gene and not a product ofconcatamerization of the R114 clone and EPI 1.

[0209] The alignment of the human EST obtained from prostate tumor withR114 revealed a very high level of identity at the 5′ and 3′ ends of theoverlapping region and a somewhat lower homology in the middle. Inaddition, 13 base insertions and deletions were identified between theEST sequence and R114. After excluding 7 of the 13 differences becausethey would have caused a frame shift, the two sequences were translatedand compared. This comparison revealed an 81% homology at the nucleicacid level and an 85% homology at the amino acid level. Interestingly,no homology was found between the two sequences before position 143 ofR114. Position 143 is six bp before the third methionine residue. Thus,the translational start site of R114 may be the third methionine residuein the ORF.

[0210] Further, 95% homology was found to exist at the nucleic acidlevel (98% at the amino acid level; there is a one base deletion in theEST that is probably an error of sequencing) between the 3′ end of theR114 ORF from position 580 to about 987 and the full length of an ESTfrom mouse mammary gland (accession # AA472513).

[0211] Northern blot analysis using a sequence from the R114 clonerevealed that the expression of the R114 mRNA was moderately upregulatedin response to the multiple MECS treatment.

[0212] Another IEG nucleic acid clone was designated R198. The followingtwo nucleic acid sequences are within the R198 clone: (SEQ ID NO:49)+TL,15′-TTTKTTTKTAATTTTTTTTTTTTNATTTGGGTTGATTCCTTGTNTTTTANTTGCCAAATNTTACCGATCANTGANCAAAGCAAGCACAGCCAAAATCGGACCTCACCTTAATTCCGTCTTCACACAATAAAAAAACGGCAAACTCACCCCCATTTTTAATTTTGTTTTTAATTTTACTTACTTATTTTATTTATTTATTTTTTGGCAAAAAAATCTCAGGAATGGCCCTGGGCCACCTACTATATTAATCATTTTGATAACATGAAAAATGATGGGCTCCTCCTAATGAAAAASCAAGGAAAGGAAAAGGCCAGGGGAATGAGCTCAAAATTGATGCCCACKTGGGGAGCATCTGGTGAATAATCGCTCACKTCTTTCTTCCACAGTACCTTGTTTTGATCATTTCCACAGCACATTTCTCCTCCARAAACSCGAAAAACACAASCGTKTGGGTTCTGCATTTTTAAGGATAARARARARAAAGAGGTTGGGTATAGTAGGACAGGTTGTCAGAAGAGATGCTGCTATGGTCACGAGGGGCCGGTTTCACCTGCTATTGTTGTCGCCTCCTTCAGTTCCACTGCCTTTATGTCCCCTCCTCTCTCTTGTTTTAGCTGTTACACATACAGTAATACCTGAATATCCAACGGTATAGTTCACAAGGGGGTAATCAATGTTAAATCTAAATAGAATTTAAAAAAAAAAGATTTTGACATAAAAGAGCCTTGATTTTAAAAAAAAAAAGAGAGAGAGATGTAATTTAAAAAGTTTATTATAAATTAAATTCAGCAAAAATTTGCTACAAAGTATAGAGAAGTATAAAATAAAAGTTATYHGTTTCAAAMTAVCDTRTCGAMCTCVTCVACCCGRGGAAKCCMCTASKKCBARHSCGGCCCCCACCSCSSYSKAKMTYCATKCTTTTGAWWCCCTTTAGTGAGGGTTAANAA-3′and (SEQ ID NO:50)5′-CAGCCTCTCACTCTCTNGCTCTCTTTCTGTCTCTTCCTCGCTCCCTCTCTTTCTCTCCTCCCTCTGCCTTCCCAGTGCATAAAGTCTCTGTCGCTCCCGGAACTTGTTGGCAATGCCTATTTTTCAGCTTTCCCCCGCGTTCTCTAAACTAACTATTTAAAGGTCTGCGGTCGCAAATGGTTTGACTAAACGTAGGATGGGACTTAAGTTGAACGGCAGATATATTTCACTGATCCTCGCGGTGCAAATAGCTTACCTGGTGCAGGCCGTGAGAGCAGCAGGCAAGTGCGATGCAGTCTTTAAGGGCTTTTCAGACTGTTTGCTCAAGCTGGGTGACAGCATGGCCAACTACCCGCAGGGCCTGGACGACAAGACGAACATCAAGACCGTGTGCACATACTGGGAGGATTTCCACAGCTGCACGGTCACAGCTCTTACGGATTGCCAGGAAGGGGCGAAAGATATGTGGGATAAACTGAGAAAAGAATCGAAAAACCTCAATATCCAAGGCAGCTTATTCGAACTCTGCGGCAGCGGCAACGGGGCGGCGGGGTCCCTGCTCCCGGCGCTTTCCGTGCTCCTGGTGTCTCTCTCGGCAGCTTTAGCGACCTGGCTTTCCTTCTGAGCACGGGGCCGGGTCCCCCCTCCGCTCACCCACCCACACTCACTCCATGCTCCCGGAAAATCGAGAGGAAAGAGCCATTCGTTCTCTAAGGACGTTGTTGATTCTCTGTTGATATTGAAACACTCATATGGGGATTGTTGGGNAAATCCTGTTTCTCTC-3′.

[0213] This clone is similar to neuretin (accession # U88958).

[0214] Another IEG nucleic acid clone was designated R233. The followingnucleic acid sequence is within the R233 clone: (SEQ ID NO:51)5′-AAACCNAGAACCCCCCTTTGNAGAACCNTTGTTTCCTTTCAAGCCCAAGGAAGGCGGGGCCCAACCTTTGGTGTTNTTTGAACAGGCCTTGAACAGGAGGNTWAGGAGAATTTCCGGTTGTGGAACCCCAACAGGAACCCCTTGGCACCCCTGGCCCCAAGGTTGTGMAACTTTGGTTTGCTTAATTTGGACCGTTTTTGCCTTGAGGATTCATGACTTTTTTTTGKGCCCTTGTGAGCCAAGATGTTGGGTTTTCCCATCAACAWTAATAACCCCTTGCTTTTTGGGGTGATTCCCCTGGGGAGTTTCCTGATGAATTCCCCCACAGCTCCTGGGGTTTTCATCTTGTTCTTACTGTTGTCTGGATTAGGAGGGCGGAGAGGGTGGACTCCCTGAGACAAGATAAGCAGGTGGAGACATAGAAGAGGGAGGGACATTTAACATAGTAACATTTTCAGAGGTGACAGAGATGATACACGGGCAGCTGGAMTTTTGTGAAGGACAGAGGAGCTGGCAGACCCACAGGGCCATACCTTTGAGGGACAGGTGAATGGCTGGTTACCAGAGACAGGACTGGTAGACAGTCAAGTACCTCACTACGATGTGCCAAGAGATYTGGGATCCTGGGAAATGTGTGGAGAAGAGGATTTGACACTCCCCACCCCCAAGGCCCTTCCCCTTTGCTGACAGCATTGCTGTGGTCGTGGCCTGTTGCCTTGTCCTCTGTCCCTGGGTGGGGCACACCCTCCTGTGCTGTGCTTGCCTTGTGCATCAATAAACCAC-3′.

[0215] This clone is similar to KIAA0273 (accession # D87463).

[0216] Another IEG nucleic acid clone was designated R24 1. The firstlibrary screen produced a clone designated R241-4. This R241-4 clonecontained a 2.0 kb fragment and a polyA tail. A second library screenusing 5′-end of R241-4 as a probe produced an additional clonedesignated R241-12. The following nucleic acid sequence is within theR241 clone: (SEQ ID NO:52)5′-GCANTTTGGAGTTATTGCTTAAAACCAGGNTAAGGCACTTTGTCCCACAGGACCCAGGATCNTAAANGGGTTGAAATTGGGNCGGGGAACCCCAGGATATAATGCNACTTTTGTTAGGGGGAGAGTTCAGCTCTAACTGGTAGTAGTGTGAAAGTAAGCACCTTGACTTCAATTTTGGAAAGCACTTGGTAAATGGAGAGAACTTTGGAGTTTCCCTATCATCTATATCAGTCTTTGAACACACCCTCAAGTCCCAGCCTCAAGGCTCAATAAAGGACCACATAGCAGGTCTGAGGCTCACTGCTCTCAGCCCTTAACACAGGGCAGTGGAGAGCAGGGTGATCTTCCCTCTCTGGAGCTTCTCCTTGGCCTTCTTCTCCACTTGGGCTTCTGCTCAGCAGCAGATATATTCTGGGTTCCATAAGGAATCCAGCTGTCCCAGTGGCTTGACCCTGTCAAGGCAAGATATCAACTCTGAGGATGACCCAGTCATGGAGGAAGAGAGTGTGACAAGATCCGCAGTTTGAAGCAAAACTGTGTTTGGTCTTTTCAAGAAACAAATGGGCACATTGAGTTCTGTTCAGTGTCAGAGGATATCTTTCCCTTTGCTCCCAGATTTCCAGAAATGGATAATGTTTTCATTTCTGTGGGAAGGGTCAAGAAACATAAAATTGCTCAACAATGCTTGCTTCCCTTGAGGGTTGTTGAGCAAAGGCCGATATGCCTCCCTGCATTCTCTTCTACCTCAAGATTTTGGAATTCAATTCTGGAACAGAAATTTATTTACACAAGAACACTTGTTGTCAGCCTTGGTTACTGTGGGAGTTACATAAGGGTGACAGTCTGTATCTTCTAARTTAAACAGGAACTGGGCTTTGGCGGCCTATTGACCCAGTTTATATCTAAATATAACTGTGGCTCCAAATGATTGGCCAATAACATTCCCTTTACCTTCAAAGTTTTCTCCATCAGTCATTTCTGTGGCAGCACAGTTCCAATGTCATATGCCCC:TGCAAATTGTGAAAGTAATTAGTGACAAAATAACCCTCCCCCCTTTCAGTGGCCAAACTGTCAGCTGTAGCAGCGCTGCGAAAGCGAGTACTACACTATGTACGGAAAG:CCTGTTCCTTATCACGGACTAGACTCAAGAAATGCCATCTCCGAACGGTGGCATTCAAGGTGGTAGTCGTTTGAATGGAACAGTCATCTATGTGGACATTGTTAAAGTGTTTTAAAGAGTATTTTGAAAATTAAGTTTACATTTTACAACTGCTTTATTTTTTATTGAAACAATTGTATATAAATATTACCCTCTTTCACTGTTAATTAAAGTAAACCTAGACCTTGTAGACAAGTGGGTCAACTGATATGTATAGAAGCTGTGATGTAGACAATACCTTTCTCTTGTGTAAATGGTCATAAATATAGCTGTTCCTGTGTTTTTATAAGTTGAGGGTATTTTGTTGTTTTATAACAACAAAATTTATTGCATTTGAAATGGTTTTTATGTAATAGAATCATGCAAACAGTGAAGGATTATAACATGGTATATGTAAATGTATAAACTTTAGAAGAAATAAATACAACAAATTTCAAAAAAAAAAAAAAAAAA-3′

[0217] Northern blot analysis using the 3′-end the R241 clone as a proberevealed the presence of two mRNA transcripts: one about 7.0-8.0 kb andthe other 4.8 kb. In addition, this analysis revealed that theexpression of the R241 mRNA was marginally upregulated in response tothe multiple MECS treatment.

[0218] Another IEG nucleic acid clone was designated R256. The firstlibrary screen produced a clone designated R256-8. This R256-8 clonecontained a 1.8 kb fragment. A second library screen using 5′-end ofR256-8 as a probe produced two additional clone designated R256-2 andR256-3. These additional clones contained each contained a 3.0 kbfragment. The following nucleic acid sequence is within the R256 clone:5′-GGCACGAGGACAGATTCTGAGA (SEQ ID NO:53)TGGAAACTTAAATTACATCCCAGAGGCAGGGAAACTATGAAGTCACCGTTCCTAGACCACCCCTTACTGAGGTTCCACGGTCACACTGACGGCAGGACCCACAAGGGCAGGGTATTGGTCTGCCCTCCTTTCTCCTGTCTGTCTGACTTACCTAACTTTGGTCTCGGCTGCTGACACTTGGAAAGGACCAAATTACTTGATAGTATTTCCCCCTGTTTGTGTAATAGCCTGAAACCTTGGAGAGGTTCCAGAATACTTCTGTATATAGGGCACAGGTGTGAAGACATTGTCCAAAGCTTATTTATTTATCTATTTATTTACCCTGGCTGAGTAACCACACCAGTAGGGGGAAAACTAAAATGTGTTGAGTGTAAACAAAGTCACCAGCCTGGCTAGAAATTCTCCCTGGAAAACATCCATTTTGATACAATGTAAACGTTAGTGTTCACCCTTAGATACATGTTGAAAGAGAGCTTTGGTACGCGGAAGTGGCATCTITGGTCACACACCATGCCAAAGTGAAGAGGTGGCCAGTGGAGGTCTTCCGGTCCTGTCGGGATCATTTGTGAATACATTCTTTGCCCCTCTTAAGTACTTGTTTACTAAACATGTGCAGTGGTAGGTATTAGTGTTAGATCACAGTGGGCACTTCCCTGGGGATCTGGGGAAGACCAGAGCTTGCAACTCTGCCTGTTTTGATCCCTATTTCTCACAGTGCTGTATTAAAAAAATAGGATTTAAGACAGATAACCACCTTTACATTGTGAGTGTGTTTGCCTTGTCTAACGACAGATAATAACCTTAACATTTCTCTTCACCTTAGTACTTTAGGCTAATTATACACGTCTGTCTATGCCATGAGTAAGTGGACTGTAGTCGGACCAAAAGAAAACAAATGAGCCGTTGGACCATTTGTGCAGTCAGTTTCTGGTCCTTAGATGTATCCTAAGCAGTAAGTGTCTGATTGTACCCTGGTGGTATGATCAGTTGTCTCGTAGCTGTCTCAGCTCCACAGTTTACAATGCAAATCTGTCTCAAGATCTTCACGTCACTGCTGCTGAGAGCAGGGAGAATTCTCTGCAGCTGTTTCAAAGTTGTGGCCCGGCCTTGAATCCTCTGTTAATTACTGTGTGAGCCAGAGGGAGCTGCCCAGCAAGGGTGGGCCCCCAGCCGGCAGGGGAACTTTCTAGACTCCCCGCTCATTCAATTGATCTAGGCATTCGGGCCTGCTACTTGACCATTCTCGCCCTGTGAAATGTCCCACACTTTGAAGCAAATACAATTCACAGCACAGTACACACAAAACCCTGGCATATAAGACAGGGGAGGTTCTTCTTATTTTGTGAGCCGGTTGCCCTGGAAACGGATAACAAAGGGCAGCCTTCCACTTCTGGCATAATGGTGGAGCCTCTTTTCTCAGGCTTGACACCTGTCTGAATAAGAGTGATTAGAGCCGCATAATATCCCTCTCTTGGCTATTGAATATGTGGTTCACATACCAAACCCTGTAGAAGTTAGAAGACGGTCGTGAACGTATGTTGTTTGCTTCCACTACATTTTTGAGGTTTTGTAAACTGTTATTTTTTTTCACGATGTGAAACTGAAGGTCAATAAATTATTAGAGATTTTCAAAAAAAAAAAAAAAAAAAAA-3′.

[0219] Northern blot analysis using a sequence from the R256 clone as aprobe revealed the presence of a 4.0-4.8 kb mRNA transcript. Inaddition, this analysis revealed that the expression of the R256 mRNAwas moderately upregulated in response to the multiple MECS treatment.

[0220] Another IEG nucleic acid clone was designated R26 1. The firstlibrary screen produced a clone containing a 1.0 kb fragment with apolyA signal and tail. A second library screen using a portion of thisclone as a probe produced 41 positive signals that were isolated. Inaddition, PCR using T3 or T7 primers along with a R261 sequence specificprimer resulted in the 850 bp of additional sequence from a solutioncontaining the phage plug from a first screen. The following nucleicacid sequence is within the R261 clone: 5′- (SEQ ID NO:54)CTTAAAACCCCTAGATTTCCTGTTACATACTAACACAGGTCTTCCCTTTCACTCCAACCCCAGGTTTCAGGCCTCAGAGCCATGCTGGGGTTGGAGAAAACTGCATTCCTATGAGGGTAAAAAGTAGCTGCCCTCTCTGACCCTTTCTTGCTAGGCTTCATGCGGGATGGGAGAGGGTATCCCCAGGATGGGGACAGAGGAAGCCTGGCTAGGGCCTTCTAGCCCAATAAGCCAAACAGGAACTATAAGCAGATCAAAATCCTACACTAGCTTATTAGGGCCCTGTTAGTTGAAAACCTTGTTGCTGTCCCAAGTTCTTCAGTTACAACCGAGTACACTTACTCTTCCAACTGTCCTAAGGGTCACTACCCAGCCAGCTTTGGATCTTCAGCACTTTTAAAAGCTGAAACTCCCTCTTGCCCTTCTTGTCTATTCCTCACTGCCAGTTGGGGCCTAGGCTCAGTCCTGGGCAAATGCCCATGATCCTGCTGCTGTGGGAAGTTTGATAGGGCATTTGGCTCAAATTTCAAAAGGCCTCGCTCCTGACCTGATTTCTCGAAGCTCCAGTAGYJCTAGACCCCTCCAATCTCTCATCTGACTGGTTGCAAGGCTTATTTTTCTTTTGTACThTCCTATAGAGCATTTCTGTAGCATTTGAGTGTGGCGATATTTTTGTTGTGTGTAGATTTCTAAGAACCAACACTACTCAGTCTCCTGCTAGTCTGACTCCTGAAGCATCAGACCTCGTCATACGGTATTGACTGTGTATGTGCCTTTCACCTTGAGCATGCTTCAGGATTTTTTTTCTTAAACCACAGAACTTGAATACACAAGGGAACCAGAATTCACAAAGTCCTATGCAACCCTAGACAGGAGGAGGTTAGAGAGTCTGTCTTGATTGGTGATTTCAGAGACCCNAGAGAAATTTGTACCAGTTTGTATTAATGTCAGTACTACCAGCACTTTGCCAAAACTAAGGATGTCAGAGGGACCTGTTTCTAGAGTGAGTCCCAATTACATCAAAGGGCAACTTACAGCTTTCTCCAGTAAGTCTGAGTGGTTCTCTTGAGCTGGTGTCACTTTCTAACCTTTGCCAGTCTAGCCCAGCAGGGCCCTGTGTGTGTGAGTGCAGTTTGGTGCTGTTTTGGAGTATGCCTGCTCCCCAGCCTGGAACCCTCTCAGCAACTTGCTGGGACCTATAATGTCTTAGGTGCAACAAGGACCCTACCAGAGCTCCTGGGTGGCTTTCAAGATCCACGTAGCTTTGTGTGAGGGGACTGAATGCAGACAAACCACAGCCTGCTTCAAATACCTTCTTTCCCTACCACCTAGTTCCAAATGGAACCAACAAGTTGAGTGCATCTCTGTTGGGTGTTTTGTGTLGAGACTGGCTGAAGTGAAAACTCTTTGACTGACCATGTTGTGATGTGTCGACAGACTCAAGGACACAACCACCTCGAGCTGGTCATGTGGCATGCCTGTGTATGTGTGTAACAGGATTCTGAATGTTAGGTTGTAATGCTATTCCTGTATGGGAGAAAAAAATAATATAAACAAATAAAAATCTATTTAAAGCACAAAAAAAAAA-3′.

[0221] Sequence analysis revealed the presence of some homology with ESTsequences including that of a cDNA clone from ae69b04.s1 Stratageneschizo brain S11 (accession # AA774320).

[0222] Northern blot analysis using a sequence from the R261 clone as aprobe revealed the presence of a 4.0 kb mRNA transcript. In addition,this analysis revealed that the expression of the R261 mRNA wasmarginally upregulated in response to the multiple MECS treatment.

[0223] Another IEG nucleic acid clone was designated R272. The firstlibrary screen produced a clone that was used in a second libraryscreen. This second library screen produced two additional clonesdesignated R272-1 and R272-2. Clone R272-1 contained a 2.0 kb fragmentwhile clone R272-2 contained a 1.7 kb fragment. The following twonucleic acid sequences are within the R272 clone:5′-CCATGGGGACTGGTTTGTCACCNATTGCCCATGGNTTTGGTT (SEQ ID NO:55)GGTAGGTGTTTTTTGGTGGACATTTTTTGTTTCNCGTTTTGAACTCCAGATTATTGGGTTTTTTGTTTTAATTTATTTTTGTCAGAGGAAAAATAATTTAACATCCATCTCACAGGCTTGCTTGACTGTTCAGTTCCAAGGTCCTGCTCACTTTTTCTTGTCTTGCCTCTGCTCTGGCTTTCTTCATGATAGTGCTGGACGTGGAGCTGAGAGTCTCGTTTACTCTAGGCAAACCCTCTACCTGAAGCCAGAGCCCAGCACTCCGTACCACCACAGACTTCTGAAGCTGGCAAAGTTTTAGAAGCTGGGAGTTTTCTGATTCTCTCATTATTAAGTTCTCCTCAGTCTTTAGATAGAGGTAAATGTGGGCTTGTAAGAAAAGAAACGAAAGCACGTAATGTACACCTATTCTGAATTATGCAAATTAGCTCTTACTCAGGGTCAACTAAATTACTTCAACTCGCCCTTTAGTTTACTCTTAATTTGCAAAAAGAGAAAAAAGAAGGAAAACTAAATAGGACTATGATTTGGGGAGCCAAATTGATAATCTGATGTAAAAGTTGCTGTGTTAAACATAAATTATTAAGTGTAGACTTTTTTCCTAGGATATTGTATTCATTTTGTGATATCGCCTAGAATGATGTATTAGATAAAAATCAATTTTGTAAGTATGTAAATATGTCATAAATAAATACTTTGACTTATTTCTCAAAAAAAAAAAAAAAAAAAAAA-3′ and5′-GATTTATATTCAATGTTGTTTATTTAATCCATTGCAGTTGGTGAATGCCTTTT (SEQ ID NO:56)CCTCCTAGACACCCTGTATTATACCATTTGGGGATTAAGTCAAAGTTAAGTATATTTTTTTCTTACTTGAGCTCTATATATGCAATTCAGATATCTTCCTGATGACAGTTTTATATGTAAATGTAATTTAACTTTCTTTCCGTGTTGACGAAGTTCTGTAGGTGTTAGGGTTAGAAGTCTCAGCACTCACTTCTCTCACTGGATGTGCAGTGTGCCTGCCATGGCGCACGGCTTCTCAGTAATGATGCCATCTCTGCTACTTTTACAGAAGGAGAATTTACTTTTGAGGTGGGTATGTGTTGATATCTAAACACTGTGTGTTGCTTGCTTAGATAGGCAAGACACACTGCTGTGCGTGGCTCCTGTGGTGCACCTAGCCCAGGGGAACGTAGCCTCAGTACTTCCGCTGGCTTCTTCATGCCTAAGAAGCAGGGGCCTTTCTTGTTTGCTGGGCTCTGGCTTTAAAAGTTGTCCTTTGGGTCTGGAGATGTAGCTCTGTGACAGAACACCAGCTAATGTCAGGTCCTGCGGTCAGTCTCTGGTACACACAAGCGCACACTCACATGATGGGGGGATGAAAGGCTGTCCTTGTGTAACAGTATTCGATGGGGCGTTGCCTGGATGACGATGTTTATGTACTCTGAAGGCAGATCCTGAAGGCACCCTGTTCTTCCCTTCCTTGTGTAACTGAGTCTGCACTAGCTTAGCCACTGTTTTAGAGGCCATCCTAGTGGGCGAACAGGAGGCATCGCACTGGGTGATGGTTTGCCTTCAGTCCTCAAGTAACAGCGGCCGACTAAATGCCGATGGCTTGTTTTTGAAATCAAATATTACCAAGTTGGCCTAGTCTGCCTTCTGTGAAGAAGGGGAGAAAGGAAGGGTGGAAAGGTGGATGGAAAGCCTTTGGGGAACTAGTCTGATCTCTCAAGGG-3′.

[0224] Northern blot analysis using a sequence from the R272 clone as aprobe revealed the presence of a 1.0 kb mRNA transcript. There appearsto be a discrepancy in the length of the R272 mRNA since the Northernblot data indicates a message of 1.0 kb while the cloning data reveals amessage length around 2.0 kb. Regardless, the Norther blot dataindicated that the R272 mRNA expression level was moderately upregulatedin response to the multiple MECS treatment.

[0225] Another IEG nucleic acid clone was designated R280. The followingnucleic acid sequence is within the R280 clone:5′-CTTCAGTTCCTTTGAGGGGNCTTTCCTTC (SEQ ID NO:51)GAAGGGGATACGCCTACCTTTCACGAGTTGCGCAGTTTGTCTGCAAGACTCTATGAGAAGCAGATAAGCGATAAGTTTGCTCAACATCTTCTCGGGCATAAGTCGGACACCATGGCATCACAGTATCGTGATGACAGAGGCAGGGAGTGGGACAAAATTGAAATCAAATAATGATTTTATTTTGACTGATAGTGACCTGTTCGTTGCAACAAATTGATAAGCAATGCTTTTTTATAATGCCAACTTAGTATAAAAAAGCTGAACGAGAAACGTAAAATGATATAAATATCAATATATTAAATTAGATTTTGCATAAAAAACAGACTACATAATACTGTAAAACACAACATATGCAGTCACTATGAATCAACTACTTAGATGGTATTAGTGACCTGTAACAGAGCATTAGCGCAAGGTGATTTTTGTCTTCTTGCGCTAATTTTTTGTCATCAAACCTGTCGCACTCCAGAGAAGCACAAAGCCTCGCAATCCAGTGCAAAGCTTGCATGCCTGCAGGTCGACTCATATGCGGTGTGAAATACCGCACAGATGCGTAAGGAGAAAATACCGCATCAGGCGGCCATCGCCCTGATAGACGGTTTTTCGCCCTTTGACGTTGGAGTCCACGTTCTTTAATAGTGGACTCTTGTTCCAAACTGGAACAACACTCAACCCTATCTCGGTCTATTC1TTTGATTTATAAGGGATTTTGCCGATTTCGGCCTATTGGTTAAAAAATGAGCTGATTTAACAAAAATTTAACGCGAATTTTAACAAAATATTAACGCTTACAATTTGCCATTCGCCATTCAGGCTGCGCAACTGTTGGGAAGGGCGATCGGTGCGGGCCTCTTCGCTATTACGCCAGCTGGCGAAAGGGGGATGTGCTGCAAGGCGATTAAGTTGGGTAACGCCAGGGTTTTCCCAGTCACGACGTTGTAAAACGACGGCCAGTGAATTGTAATACGACTCACTATAGGGCGAATTGGGTACCGGGCCCCCCCTCGAGGTCGACGGTATCGATAAGCTTGATATCGAATTCGGCACGAGCCGCAGCCGATATGCAGTCCCCGGCGGTGCTCGTCACCTCCAGGCAAGTTCAGAATGCGCACACGGGYCTCGACCTGACTGTACCACAGCACCAGGAGGTGCGGGGTAAGATGATGTCAGGCCATGTGGAGTACCAGATCCTGGTGGTGACCCGGTTGGCTGTGTTCAAGTCAGCCAAGCACCGGCCCGAGGATGTCGTCCAGTTCTTGGTCTCCAAAAAATACAGCGAGATCGAGGAGTTTTACCAGAAACTGTACAGTCGTTACCCAGAAGCCAGCCTGCCCCCACTGCCTAGGAAGGTCCTGTTTGTCGGGGAGTCTGACATCCGGGAAAGGAGAGCCATGThTGATGAGATTCTACGCTGTGTCTCCAAGGATGCCCAGTTGGCGGGCAGCCCAGAGCTGCTAGAATTCTTAGGCACCAGGTCCCCGGGGGCTACAGGCTTTGCCACCCGAGATCCCTCTGTCTTGGGATGACGACAGCCAGCCGGCCAGGGGACAGTGATGAGGCTTTTGACTTCTTTGAGCAACAGGGATGAAGTGCAAGCCACCCACATTGGGCCTGAGCAACAANGAAATGTTGAGAAGGTCCNTGGAAGGAANGAGGAGGGAAGGGAGGAAGGANGATAACTTGGGATCCCCCTTGGGGCAATCAATGCGGCCTCCCAAAGGAAAGNCCCTAAAG-3′.

[0226] Northern blot analysis using a sequence from the R280 clonerevealed that the expression of the R280 mRNA was upregulated inresponse to the multiple MECS treatment.

[0227] Another IEG nucleic acid clone was designated R286. The firstlibrary screen produced a clone that was used as a probe for a secondlibrary screening. Briefly, the ³²P-labeled probe was used to screen aUniZAP rat hippocampal oligo(dT) primed library (Stratagene). Thissecond screening produced a clone having a 4.7 kb full-length R286 cDNAsequence. The nucleic acid sequence of this rat version of R286 is asfollows: 5′- (SEQ ID NO:58) CTGCCAGCCGAGGCTCCTGCCGCTGTGACCCGCGCTCCGCCCGCCGCCGGGCCGGGACCCTGATAGCTAATGTCAGAAGAAAGTGACTCTGTGAGAACCAGCCCCTCTGTGGCCTCACTCTCCGAAAATGAGCTGCCACCGCCTCCCCCGGAACCTCCCRGCTACGTGTGCTCGCTGACAGAAGACTTGGTCACCAAGGCCAGGGAAGAGCTTCAGGAGAAGCCCGAGTGGAGACTCCGGGATGTGCAGGCCCTTCGAGACATGGTACGGAAGGAGTACCCATACCTGAGTACATCGCTGGATGATGCCTTCCTGTTGCGCTTTCTGAGGGCCCGAAAGTTTGATTATGACCGGGCCCTGCAGCTGCTGGTCAACTACCATGGCTGCAGGCGGAGCTGGCCAGAGGTCTTCAGCAACCTGAGGCCATCAGCCCTGAAAGACGTTCTTAACTCTGGATTCCTCACAGTGCTGCCCCACACAGACCCCAGGGGCTGCCATGTCCTCTGCATCCGACCAGACAGATGGATACCGAGCAACTACCCGATCACCGAGAACATCCGCGCCATCTACTTGACGTTAGAAAAACTCATTCAGTCCGAGGAGACCCAGGTGAACGGGGTTGTAATCCTCGCCGACTACAAGGGAGTGAGCTTATCAAAGGCGTCTCACTTTGGCCCCTTTATCGCCAGAAAGGTGATTGGCATCCTTCAGGATGGCTTCCCCATTCGGATAAAAGCAGTTCACATAGTAAACGAACCTCGGATATTTAAGGGCATTTTCGCCATCATAAAACCATTTCTGAAGGAGAAAATTGCAAACAGGTTCTTCCTCCATGGGTCTGACCTGAGCTCTCTGCACACGAGCCTTCCAAGGAATATCCTCCCCAAAGAGTATGGGGGCACCGCTGGGGAGCTGGACACTGCCAGCTGGAACGCGGTGCTGCTGGCCTCGGAGGATGATTTTGTGAAAGAGTTCTGCCAGCCTGAGTCTGGCTGCGATGGTCTCTTGGGCCAGCCCCTGCTGCCTGAGGGGCTGATCTCAGACGCGCAGTGTGACGACTCCATGCGAGCCATGAAGTCCCAGCTCTACTCCTGCTATTAGCCCTCTTCCGGGAGAATCACCATGTGTAATTCCTTCCTTCTTCGAATGCACAGGCTGAAGATGCCAGGACCTCGGTCTTGCTCCATCACAGTGCAGCACGGAGCTGCCTGCAGAGATTTAAGGAGAGCCCATCACAGGCAGACCTCTGACCAGCTAGGTTATTCCAAGAAGACATGGAAATTGCCCTGGTGATTCCCAGATGTCTGTACTCTAAGTCTGCAACTGTTACTCTGGAAGCTGCATCTGTTTCTTATGCATCTTGGAAAGAACTAGGGTCAAAGTCACTCTGAAGTGACCAGGAGTAGACAACTTGATTGATCATGAGTCTGAAACAATTGCCAATCCTGAAAGGTGGCCATGCGTGAGACTTTGAGTCTCTTTCCCATAAACTGTAGGTGTTGACTACTGCTGCTTATCTGCAAAGGTCAGGGTTCAGGCCCCAGTTGGCATTGCTGGGTCTGGGAAGCACTGCTAACTGAGTGGTAGAAACGCCAGGCCCAGGCAGCACTTAAAGGTTAAAGGTCAAATTTGGAAGCTAAGGCTATAAATCATCCTGGGTTCCAGGCTTAAATCTTGCAATGGACACTCTCCCCAAACCATAAAGCCTTAGCTCTGGTTCTCCATGGAATCATGCAGGTCAACATAAAATACTGGATTCTTGGACTGCGTGGCTAAAAGCACTTAGACTARGAGTCCAGTGTGTGACTGGATGGATAGGGGCCTCAGCTTGTCAACTCTAAGTTAGMGMTCCATGGAATGAAGGCCTTGRGGGCTGCTCAAGTTCTGYLAGGTTTCTGCTTTGGAAAGATGACCACCTGGAGGTGGCCGGGCCTTTTTGGTTTGGCTTGGTTTTGTGTTATAGACACAAGCCTTTATGGAAAGGAACCGTCTGGCCTTTAAAGAAATTACTATGTTCCTGGGAGTTGGTGGTAACCAGCTGCTTTTGCAGATGATGGGTGAACTGGAAAGGGATGGCTTTTGTGAGGCTGACCAAGTCTTGTACGCGGATGTTGTACAGATTTCCTCCCACACCGGAGACATTCGTACTATATTAGAAACAGCCACGGACTTGTGCTCTTTTTCAGTTTGTGTCCCTGGAAACATACGGGGGGCAGGCTGTTGCTGGTTLCACCTGGGGGCCCTGCCCTCCCAGACACGGGAGTGCTTGTCTAGCGTGGGAGGGCCAGTTGGCCAGATTGTTAGCTCTGCGTTGGGGTGTCGTAGACAACTGACAGGATTTTAGCCTTAACCCAAGCACTGAGTGAGGTGATTTTTCCCTTGGCTTTTGGCGTGTCTTTGGTATTCACCATGTATTGTGGTGTCAGGTAGTGTCAGGTACTGTTGGCTGTGTGTCTCCTAGACTAAGCGGGCGTTGSATACAGCTTACATACAGTGCTTGGAGACCAAAGGTCAGTTGGTTGTAATAAGCTGGTCCACCCTTAACAGACTTCCCAAACATYACAGAAGCTYTLATGGMCCYLACCTAATAATGCCAATTCTGGAGGACACTCTTTTACCATAGAWKCSAATCCTTTGATCTCCTGGCTCCTGGTTGAGCTTCCGCACTGATACACCCTCTTGRCTGCCCATCAGGGCCATTTGCTGCTGAGTTCTGCATTGCTTAAKCTSCKGSYGYYLTCTGCCTAAAGGGATGGCCACCCAGACACCTAAAAAGACCCGGGATGGCTCTCTAGCCTTGGTGGAGAGTCTTATTAGAAGTTTTCTTTGGGGGATTGGGGATTTGGCTCAGTGGTAGAGCGCTTGCCTGGCAAGCACAAGGCCCTGGGTTCGGTCCCCAGCTCTTAAAAAAAAAAAAAGTTTTCTTTGGTAGTTGGGGAAAAGGCAGAAGGAAAAAAACAAAGGGAAAGATGAATCTCTCAGTCCTACCTGGTTCCCTAAATTAAATCGTGTCATGTGACTAGTTAAGTCTCTTTGACTTTAACAAAGGGACACCAGGTTCTTGGGGAGAAATCTCAGAGCAAAATGTTGCCTGTTGSTAACCTTCTGGTAACCARAGGARCCTTGATAARCTTARGAGYKGACTGTATGTCCATGCTCTTGTGACTCTAGAGACTCTGGCACCTCAGGTTNAAGCAGGCTGTGAGCCAGATGTCCTGGTGCCAAGCAACCCCACTGTTGAGCAGCAGGGGCACCATAGGCCTCAGCTAGGGGAGCGCACTGGTAGAGCCAGCAAGTGAGCAGGAATCTGACTTTAGGGTAAAAATCTAGACAGTTCTGACAGCTGGAAGTCAACTTTTCCTCCATTCAAAGTCATGTGGCATTGGGAAGGGGCTAGGGAAATAGAAGTGGGTTCCAGCTTTATCTTCCTACACAGTCTCGAGTATAGCATTAACACCGAGTGCTGGACAGAGGTTGTCTGCTGAACACTCAATCCTGCTCCTGACTGACTCTGGAAATAAGGACAYLCCACTCTGCTTGGCGCGGAGATGCCCTAGTGTGCGGCCGCGGGGGCTLCTCTTTCTCAAGTCCTCTACAGNACTTCCAGGCAGTTCATCTTCCTAGGAAAAGGTATGGAGGTTCTGCCTTCATGGTAGAAACACAGGATAAAATCTACAGTAAACAACCGGTAAGTGCTGGCTTCTTACGCCTTGGCTTTCTCCAGGCACAGGTGGGTTCGACTACTCCCATTTCATCTTTGTAAGCACCTCAGGTTATAGGGCAGTTTCTTCAGAGTTGGGGGGACTGGAGCCATTCCCCCTGTAATGCCTGAGGTGGCCTTACCACCTAGCAGCCAGTTTGGCCAGCAACAGCCACACTGCTG1TATGGTATCATAATACCTCATCCTCGGGTTTCCTTCAGAAAGGRAAAWGCTAACTCAGTTGATGTAAGTGTTGCTGTGCTGGGATCCTGTCATGTGGGAGGGAACACCAAATACACAGGCTCTCAGGAGACATCTTGCTAAGGCTTCTCTTTACTGCAGTCTGCTCACGTTGTAAATCTGCCCTCTGTTCTCCTGACTCARAAAGACTCAGCCMCAAATCAAGAAGCGCCATCAAACGTTCCTTCTCAKKGGGAACGTGCTCCACAGGAAGGTCCAGWGGGATTTGCARCTAGAGTCACGTTTTACTGGKTTGTGAMCAAATTTACTGGTTTTCARTTACCTGGGGKCCTATGKGKKTTTTMAACCTTTTCCCATMAGGCAGTTAGTAGTAGCCACTTTGGGTTCCTGTGGACGTGCCTCAGCTTCTCGGCATAGGAACCCAACAGGTAGAATACTTGAAACTTCTCAGTGGCCAAGACCTCGATACCCTCTCTGATGGGTGGGAACTGGGCTATTTTCCTGACCAATCTAGGCCACCATTTTAGTCCCTGGTCACATTCCTTACTCCAAACTGAAATTCAGTTTGGCTTTGAGTATGTGCACACGTGGTGGGTTCACCTACTTCAGTGTTGACCAAAAGTTTATTTTTCTAGTGCATTTTTCTAAATGGTAAAAATATGTAATTTTAGTATGCATGACTGGGTCTCCAAAATAAAAACTGAGTGTATTGTGAAAAAAAAAAAAAAAAAAAAAAA AAAA-3′.

[0228] The following nucleic acid sequence is the ORF for rat R286: 5-′(SEQ ID NO:59) ATGTCAGAAGAAAGTGACTCTGTGAGAACCAGCCCCTCTGTGGCCTCACTCTCCGAAAATGAGCTGCCACCGCCTCCCCCGGAACCTCCCGGCTACGTGTGCTCGCTGACAGAAGACTTGGTCACCAAGGCCAGGGAAGAGCTTCAGGAGAAGCCCGAGTGGAGACTCCGGGATGTGCAGGCCCTTCGAGACATGGTACGGAAGGAGTACCCATACCTGAGTACATCGCTGGATGATGCCTTCCTGTTGCGCTTTCTGAGGGCCCGAAAGTTTGATTATGACCGGGCCCTGCAGCTGCTGGTCAACTACCATGGCTGCAGGCGGAGCTGGCCAGAGGTCTTCAGCAACCTGAGGCCATCAGCCCTGAAAGACGTTCTTAACTCTGGATTCCTCACAGTGCTGCCCCACACAGACCCCAGGGGCTGCCATGTCCTCTGCATCCGACCAGACAGATGGATACCGAGCAACTACCCGATCACCGAGAACATCCGCGCCATCTACTTGACGTTAGAAAAACTCATTCAGTCCGAGGAGACCCAGGTGAACGGGGTTGTAATCCTCGCCGACTACAAGGGAGTGAGCTTATCAAAGGCGTCTCACTTTGGCCCCTTTATCGCCAGAAAGGTGATTGGCATCCTTCAGGATGGCTTCCCCATTCGGATAAAAGCAGTTCACATAGTAAACGAACCTCGGATATTTAAGGGCATTTTCGCCATCATAAAACCATTTCTGAAGGAGAAAATTGCAAACAGGTTCTTCCTCCATGGGTCTGACCTGAGCTCTCTGCACACGAGCCTTCCAAGGAATATCCTCCCCAAAGAGTATGGGGGCACCGCTGGGGAGCTGGACACTGCCAGCTGGAACGCGGTGCTGCTGGCCTCGGAGGATGATTTTGTGAAAGAGTTCTGCCAGCCTGAGTCTGGCTGCGATGGTCTCTTGGGCCAGCCCCTGCTGCCTGAGGGGCTGATCTCAGACGCGCAGTGTGACGACTCCATGCGAGCCATGAAGTCCCAGCTCTACTCCTGCTATTAG-3′

[0229] Using the rat R286 cDNA sequence and a portion of the human R286nucleic acid sequence, specific primers were designed to amplify thehuman R286 homologue. After RT-PCR using human hippocampal RNA and thespecific primers, the PCR product was subcloned in the TA-cloning vector(InVitrogen) and sequenced with SP6 and T7 primers. The followingnucleic acid sequence is the ORF for human R286: 5-′ (SEQ ID NO:60)ATGTCCGAAGAAAGGGACTCTCTGAGAACCAGCCCTTCTGTGGCCTCACTCTCTGAAAATGAGCTGCCACCACCACCTGAGCCTCCGGGCTATGTGTGCTCACTGACAGAAGACCTGGTCACCAAAGCCCGGGAAGAGCTGCAGGAAAAGCCGGAATGGAGACTTCGAGATGTGCAGGCCCTTCGTGACATGGTGCGGAAGGAGTACCCCAACCTGAGCACATCCCTCGACGATGCCTTCCTGCTGCGCTTCCTCCGAGCCCGCAAGTTTGATTACGACCGGGCCCTGCAGCTCCTCGTCAACTACCACAGCTGTAGAAGAAGCTGGCCCGAAGTCTTCAATAACTTGAAGCCATCAGCCTTAAAAGATGTCCTTGCTTCCGGGTTCCTCACCGTGCTGCCCCACACTGACCCCAGGGGCTGCCATGTCGTCTGCATCCGCCCAGACAGATGGATACCAAGCAACTATCCAATTACTGAAAACATCCGAGCCATATACTTGACCTTAGAAAAACTCATTCAGTCTGAAGAAACCCAGGTGAATGGAATTGTAATICTTGCAGACTACAAAGGAGTGAGTTTATCAAAAGCATCTCACTTTGGCCCTTTTATAGCCAAAAAGGTGATTGGCATCCTCCAGGATGGTTJCCCCATTCGGATAAAAGCAGTCCATGTGGTGAATGAACCTCGAATATTTAAAGGCATTTTTGCCATCATAAAACCATTTCTAAAGGAGAAAATAGCAAACAGATTCTTCCTCCATGGGTCTGACTTGAACTCTCTCCACACAAACCTTCCAAGAAGCATCCTCCCCAAGGAGTATGGGGGCACGGCTGGGGAGCTGGACACTGCCACCTGGAACGCAGTACTGCTGGCTTCAGAAGACGATTTTGTGAAAGAGTTCTGCCAACCTGTTCCTGCCTGTGACAGCATCCTGGGCCAGACGCTGCTGCCCGAGGGCCTGACCTCAGATGCACAGTGTGACGACTCCTTGCGAGCTGTGAAGTCACAGCTGTACTCCTGCTA CTAG-3′

[0230] The R286 clones were found to be homologous to a family oftransfer proteins for hydrophobic ligands (such as lipid solublevitamins and phospholipids). Thus, R286 is a lipid transfer polypeptide.The amino acid sequence of the rat R286 polypeptideMSEESDSVRTSPSVASLSENELPPPPPEPPXTTCSLTEDLVTKAREEL (SEQ ID NO:61)QEKPEWRLRDVQALRDMVRIKEYPYLSTSLDDATLLRTLRARKTDYDRALQLLVNYHGCRRSWPEVTSNLRPSALKDVLNSGTLTVLPHTDPRGCHVLCIRPDRWIPSNYPLTENIRAIYLTLEKLIQSEETQVNGVVILADYKGVSLSKASHTGPFIARKVJGJLQDGTPLRJKAVHIVNEPRTTKGTTAIIKPTLKEKJANRTTLHGSDLSSLHTSLPRMLPKEYGGTAGELDTASWNAVLLASEDDTVKETCQPESGCDGLLGQPLLPEGLISDAQCDDSMRAMKSQLYSCY. The amino acidsequence of the human R286 polypeptide is as follows: MSEERDSL (SEQ IDNO:62) RTSPSVASLSENELPPPPEPPGTTCSLTEDLVTKAREELQEKPEWRLRDVQALRDMVRKEYPNLSTSLDDATLLRTLRARKTDYDRALQLLVNYLISCRRSWPEVTNNLKPSALKDVLASGFLTVLPHTDPRGCHVVCIRPDRWIPSNYPTTENLRAIYLTLEKLJQSEETQVNGIVILADYKGVSLSKASHFGPFIAKKVIGILQDGFPIRIKAVHVVNEPRTTKGTTATTKPTLKEKIANRFFLHGSDLNSLHTNLPRSJLPKEYGGTAGELDTATWNAVLLASEDDFVKETCQPVPACDSIGLQTLLPEGLTSDAQCDDSLRAVKSQLYSCY.

[0231] Northern blot and in situ analysis using a sequence from the R286clone as a probe revealed the presence R286 mRNA throughout rat brain.For in situ hybridization, Dig-labeled cRNA probes were used asdescribed elsewhere (Kuner et al., Science 283:5398 (1999)).Specifically, R286 mRNA expression was the highest in the cortex andhippocampus while being moderately high in the cerebellar granule cells,brainstem nuclei, several lateral and medial thalamic nuclei, olfactorybulb, and striatum. In addition, this analysis revealed that theexpression of the R286 mRNA was upregulated in response to the multipleMECS treatment. Briefly, a probe from the 3′ untranslated region of R286was used to hybridize a Northern blot containing 2 μg polyA⁺ RNA fromhippocampus from brains of untreated rats as well as rats receiving themultiple MECS treatment. After one day of exposure using thephosphoimager FLA2000 (Fuji), an upregulation of R286 mRNA was detectedin the hippocampus (3.72 fold induction) collected four hours after thelast MECS treatment. An additional Northern blot analysis using 10 μgtotal RNA from hippocampus from untreated rats and rats receiving themultiple MECS treatment was performed. In this experiment, the probe wasthe ORF of R286 and the level of expression was found to be induced 2.4fold in the MECS treated animals (Table I).

[0232] In addition, rats that developed seizures followingintraperitoneal injection of kainate or PTZ were analyzed for theexpression of R286 mRNA in addition to the mRNA of other IEG clones(Tables III and V). R286 mRNA expression was observed, by in situhybridization, to be mildly upregulated in the hippocampal pyramidalcell layer, cortex, thalamus, and cerebellar Purkinje cell layer at 6hours post-kainate injection. At 6 hours post-PZT injection, R286 mRNAexpression was observed to be mildly upregulated in these brainstructures, while no upregulation was observed at 20 minutes post-PTZinjection or at 1.5 hours post-kainate injection.

[0233] Other IEG nucleic acid clones included L073 (concatamer withKrox-20), L125 (oxoglutarate carrier protein), L201 (concatamer), R094(fra2), and R217 (diacylglycerol kinase; accession #D78588).

OTHER EMBODIMENTS

[0234] It is to be understood that while the invention has beendescribed in conjunction with the detailed description thereof, theforegoing description is intended to illustrate and not limit the scopeof the invention, which is defined by the scope of the appended claims.Other aspects, advantages, and modifications are within the scope of thefollowing claims.

What is claimed is:
 1. An isolated nucleic acid comprising at least oneadenine base, at least one guanine base, at least one cytosine base, andat least one thymine or uracil base, wherein said isolated nucleic acidis at least 12 bases in length, and hybridizes to the sense or antisensestrand of a second nucleic acid under hybridization conditions, saidsecond nucleic acid having a sequence as set forth in SEQ ID NO: 1, 2,5, 6, 7, 8, 9, 10, 13, 14, 15, 16, 17, 20, 22, 23, 24, 25, 28, 29, 33,34, 35, 37, 39, 40, 41, 42, 43, 44, 47, 49, 50, 51, 52, 53, 54, 55, 56,or
 57. 2. The isolated nucleic acid of claim 1, wherein saidhybridization conditions are moderately stringent hybridizationconditions.
 3. The isolated nucleic acid of claim 1, wherein saidhybridization conditions are highly stringent hybridization conditions.4. An isolated nucleic acid, wherein said isolated nucleic acidcomprises a nucleic acid sequence that encodes an amino acid sequence atleast five amino acids in length, said amino acid sequence comprising atleast three different amino acid residues, and being identical to acontiguous portion of sequence set forth in SEQ ID NO: 11, 21, 30, 36,38, or
 48. 5. An isolated nucleic acid comprising a nucleic acidsequence at least 60 percent identical to the sequence set forth in SEQID NO: 1, 2, 5, 6, 7, 8, 9, 10, 13, 14, 15, 16, 17, 20, 22, 23, 24, 25,28, 29, 33, 34, 35, 37, 39, 40, 41, 42, 43, 44, 47, 49, 50, 51, 52, 53,54, 55, 56, or
 57. 6. An isolated nucleic acid, wherein said isolatednucleic acid comprises a nucleic acid sequence that encodes an aminoacid sequence at least 60 percent identical to the sequence set forth inSEQ ID NO: 11, 21, 30, 36, 38, or48.
 7. An isolated nucleic acidcomprising a nucleic acid sequence as set forth in SEQ ID NO: 1, 2, 5,6, 7, 8, 9, 10, 13, 14, 15, 16, 17, 20 ,22, 23, 24, 25, 28, 29, 33, 34,35, 37, 39, 40, 41, 42, 43, 44, 47, 49, 50, 51, 52, 53, 54, 55, 56, or57.
 8. A substantially pure polypeptide comprising an amino acidsequence encoded by a nucleic acid of claim
 1. 9. A substantially purepolypeptide comprising an amino acid sequence as set forth in SEQ ID NO:11, 21, 30, 36, 38, or
 48. 10. A substantially pure polypeptidecomprising an amino acid sequence at least 60 percent identical to thesequence set forth in SEQ ID NO: 11, 21, 30, 36, 38, or
 48. 11. Asubstantially pure polypeptide comprising an amino acid sequence atleast five amino acids in length, said amino acid sequence comprising atleast three different amino acid residues, and being identical to acontiguous stretch of sequence set forth in SEQ ID NO: 11, 21, 30, 36,38, or
 48. 12. A host cell containing an isolated nucleic acid ofclaim
 1. 13. The host cell of claim 12, wherein said host cell is aeukaryotic cell.
 14. An antibody having specific binding affinity for anamino acid sequence encoded by a nucleic acid of claim
 1. 15. Theantibody of claim 14, wherein said antibody is monoclonal.
 16. Theantibody of claim 14, wherein said antibody is polyclonal.
 17. A cDNAlibrary comprising a plurality of clones, wherein each clone comprises acDNA insert and wherein at least about 15 percent of said clonescomprise cDNA derived from immediate early genes.
 18. The cDNA libraryof claim 17, wherein at least about 20 percent of said clones comprisecDNA derived from immediate early genes.
 19. The cDNA library of claim17, wherein at least about 25 percent of said clones comprise cDNAderived from immediate early genes.
 20. The cDNA library of claim 17,wherein said immediate early genes are immediate early it genesresponsive to a maximal electroconvulsive seizure.
 21. The cDNA libraryof claim 17, wherein said cDNA library is a subtracted cDNA library. 22.The cDNA library of claim 2 1, wherein said subtracted cDNA library isIEG-Reg cDNA library.
 23. The cDNA library of claim 21, wherein saidsubtracted cDNA library is IEG-Lg cDNA library.
 24. An isolated nucleicacid derived from a cDNA library, wherein said cDNA library comprises aplurality of clones, wherein each clone comprises a cDNA insert andwherein at least about 15 percent of said clones comprise cDNA derivedfrom immediate early genes.
 25. The isolated nucleic acid of claim 24,wherein said isolated nucleic acid comprises a nucleic acid sequence ofan immediate early gene.
 26. A method of obtaining immediate early genenucleic acid, said method comprising: a) providing a cDNA library, saidcDNA library comprising a plurality of clones, wherein each clonecomprises a cDNA insert and wherein at least about 15 percent of saidclones comprise cDNA derived from immediate early genes; b) contactingat least a portion of said cDNA library with a probe, said probecontaining at least one nucleic acid having a nucleic acid sequencederived from an immediate early gene; and c) selecting a member of saidplurality of clones based on the hybridization of said at least onenucleic acid to said member under hybridization conditions, said membercomprising said immediate early gene nucleic acid.
 27. A method oftreating an animal having a deficiency in a neuron's immediate earlygene responsiveness to a stimulus, said method comprising administeringa nucleic acid of claim 1 to said animal such that the effect of saiddeficiency is minimized.
 28. The method of claim 27, wherein saiddeficiency comprises a reduced level of expression of an immediate earlygene.
 29. The method of claim 27, wherein said stimulus influenceslearning or memory.
 30. The method of claim 29, wherein said stimuluscomprises a maximal electroconvulsive seizure.
 31. A method of treatingan animal having a deficiency in a neuron's immediate early generesponsiveness to a stimulus, said method comprising administering atherapeutically effective amount of a substantially pure polypeptide ofclaim 8 to said animal such that the effect of said deficiency isminimized.
 32. A method of treating an animal having a deficiency in aneuron's immediate early gene responsiveness to a stimulus, said methodcomprising administering an effective amount of cells to said animalsuch that the effect of said deficiency is minimized, said cellscontaining a nucleic acid of claim
 1. 33. A method of treating an animalhaving a deficiency in a neuron's immediate early gene responsiveness toa stimulus, said method comprising administering a therapeuticallyeffective of antibodies to said animal such that the effect of saiddeficiency is minimized, said antibodies having specific bindingaffinity for an amino acid sequence encoded by a nucleic acid ofclaim
 1. 34. The method of claim 33, wherein said deficiency comprisesan elevated level of expression of an immediate early gene.
 35. A methodof identifying a compound that modulates immediate early geneexpression, said method comprising: a) contacting a test compound withan immediate early gene nucleic acid; and b) determining whether saidtest compound effects the expression of said immediate early genenucleic acid, wherein the presence of an effect indicates that said testcompound is said compound.
 36. The method of claim 35, wherein saidimmediate early gene nucleic acid comprises a nucleic acid sequence asset forth in SEQ ID NO: 1, 2, 5, 6, 7, 8, 9, 10, 13, 14, 15, 16, 17, 20,22, 23, 24, 25, 28, 29, 33, 34, 35, 37, 39, 40, 41, 42, 43, 44, 47, 49,50, 51, 52, 53, 54, 55, 56, or
 57. 37. The method of claim 35, whereinsaid effect is a reduction in the expression of said immediate earlygene nucleic acid.
 38. The method of claim 35, wherein said effect is anincrease in the expression of said immediate early gene nucleic acid.39. A method of identifying a compound that modulates immediate earlygene polypeptide activity, said method comprising: a) contacting a testcompound with an immediate early gene polypeptide; and b) determiningwhether said test compound effects the activity of said immediate earlygene polypeptide, wherein the presence of an effect indicates that saidtest compound is said compound.
 40. The method of claim 39, wherein saidimmediate early gene polypeptide comprises an amino acid sequenceencoded by a nucleic acid of claim
 1. 41. The method of claim 39,wherein said immediate early gene polypeptide comprises an amino acidsequence as set forth in SEQ ID NO: 11, 21, 30, 36, 38, or
 48. 42. Themethod of claim 39, wherein said effect is a reduction in the activityof said immediate early gene polypeptide.
 43. The method of claim 39,wherein said effect is an increase in the activity of said immediateearly gene polypeptide.